WO2021238890A1 - 一株鼠李糖乳杆菌及其在抑制幽门螺杆菌中的应用 - Google Patents

一株鼠李糖乳杆菌及其在抑制幽门螺杆菌中的应用 Download PDF

Info

Publication number
WO2021238890A1
WO2021238890A1 PCT/CN2021/095678 CN2021095678W WO2021238890A1 WO 2021238890 A1 WO2021238890 A1 WO 2021238890A1 CN 2021095678 W CN2021095678 W CN 2021095678W WO 2021238890 A1 WO2021238890 A1 WO 2021238890A1
Authority
WO
WIPO (PCT)
Prior art keywords
helicobacter pylori
lactobacillus rhamnosus
product
ccfm1119
preventing
Prior art date
Application number
PCT/CN2021/095678
Other languages
English (en)
French (fr)
Inventor
翟齐啸
陈卫
赵建新
张美怡
崔树茂
陆文伟
田丰伟
于雷雷
王刚
张灏
Original Assignee
江南大学
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 江南大学 filed Critical 江南大学
Publication of WO2021238890A1 publication Critical patent/WO2021238890A1/zh
Priority to US17/711,257 priority Critical patent/US20220228106A1/en

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
    • A23C9/1234—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt characterised by using a Lactobacillus sp. other than Lactobacillus Bulgaricus, including Bificlobacterium sp.
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/38—Other non-alcoholic beverages
    • A23L2/382—Other non-alcoholic beverages fermented
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/40—Complete food formulations for specific consumer groups or specific purposes, e.g. infant formula
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/175—Rhamnosus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus

Definitions

  • the invention relates to a strain of Lactobacillus rhamnosus and its application in inhibiting Helicobacter pylori, belonging to the field of microbial technology and medical technology.
  • Helicobacter pylori is a microaerophilic gram-negative bacterium that colonizes the surface of the gastric mucosa and the duodenum. It was first discovered by Marshall and Warren, and the two scholars won the 2005 Nobel Prize in Physiology or Medicine for this. Helicobacter pylori is one of the most important epidemic pathogens, and its detection rate in the global population has exceeded 50%. The low level of economic development and poor sanitary conditions are the two main factors for the prevalence of Helicobacter infection. In developed countries, the infection rate of Helicobacter pylori in adults is about 30-40%, and in developing countries, the infection rate of Helicobacter pylori in adults is as high as about 80-90%.
  • Helicobacter pylori infection is generally difficult to clear spontaneously and lead to lifelong infections. Unless eradication treatment is performed, or the bacteria are difficult to colonize when severe intestinal metaplasia occurs in the gastric mucosa, Helicobacter pylori will disappear automatically. Studies have shown that long-term Helicobacter pylori infection can cause chronic gastritis and duodenal ulcer, and eventually develop into gastric cancer.
  • Gastrointestinal Symptom Rating Scale is an important indicator for evaluating gastric health symptoms. It includes 15 symptoms such as abdominal pain, acid reflux, nausea and vomiting, abdominal ringing, belching, increased defecation, and incomplete defecation.
  • triple or quadruple therapy combined with antibiotics is used to eliminate Helicobacter pylori in patients to restore gastrointestinal symptoms caused by Helicobacter pylori infection.
  • the drug resistance of Helicobacter pylori is likely to increase, and in the process of treating patients with the above treatment methods, patients often have serious adverse reactions (such as abdominal pain, nausea, diarrhea, etc.). As a result, the effective rate of treatment is reduced, and the treatment effect is often not up to expectations.
  • the technical problem to be solved by the present invention is to provide a strain of Lactobacillus rhamnosus that can inhibit Helicobacter pylori.
  • the present invention provides a strain of Lactobacillus rhamnosus CCFM1119.
  • the Lactobacillus rhamnosus CCFM1119 is deposited in the Guangdong Provincial Microbial Culture Collection, the deposit number is GDMCC No: 61013, and the date of preservation It is May 06, 2020.
  • the Lactobacillus rhamnosus CCFM1119 was derived from fresh stool samples of healthy people in Kunshan City, Jiangsu province.
  • the 16S rDNA sequence of this strain was sequenced and analyzed as shown in SEQ ID NO. 1.
  • the sequence obtained was nucleic acid in GeneBank Sequence alignment showed that the strain was Lactobacillus rhamnosus, named Lactobacillus rhamnosus CCFM1119.
  • the colony of the Lactobacillus rhamnosus CCFM1119 on the MRS solid medium is milky white semi-circular convex, smooth and moist surface, and neat edges.
  • the present invention also provides the application of the Lactobacillus rhamnosus CCFM1119 in inhibiting Helicobacter pylori without the purpose of disease diagnosis and treatment.
  • the present invention also provides a Helicobacter pylori inhibitor, which contains the aforementioned Lactobacillus rhamnosus CCFM1119.
  • the invention also provides the application of the Lactobacillus rhamnosus CCFM1119 in the preparation of products for preventing and/or treating Helicobacter pylori infection.
  • the number of viable cells of the Lactobacillus rhamnosus CCFM1119 is not less than 5 ⁇ 10 9 CFU/mL or 5 ⁇ 10 9 CFU/g.
  • the product includes food or medicine.
  • the medicine contains the aforementioned Lactobacillus rhamnosus CCFM1119, a drug carrier and/or a pharmaceutical excipient.
  • the drug carrier comprises microcapsules, microspheres, nanoparticles and/or liposomes.
  • the pharmaceutical excipients include excipients and/or additives.
  • the excipients include binders, fillers, disintegrants and/or lubricants.
  • the additive includes a solubilizer, a co-solvent, a latent solvent, and/or a preservative.
  • the dosage form of the medicine is powder, granule, capsule, tablet, pill or oral liquid.
  • the food is a health food; or the food is a dairy product, a bean product, or a fruit and vegetable product produced using a starter containing the above-mentioned Lactobacillus rhamnosus CCFM1119; or the food is Beverages or snacks containing the above-mentioned Lactobacillus rhamnosus CCFM1119.
  • the preparation method of the starter is to inoculate the above-mentioned Lactobacillus rhamnosus CCFM1119 into the culture medium at an inoculum of 2 to 4% of the total mass of the culture medium, and cultivate it at 37°C for 18 hours , Obtain the culture solution; centrifuge the culture solution to obtain the bacteria; wash the bacteria with physiological saline for 3 times and then resuspend with the freeze-dried protective agent to obtain the resuspension; the resuspension is lyophilized by the vacuum freezing method to obtain Starter.
  • the mass ratio of the freeze-dried protective agent to the bacterial cells is 2:1.
  • the freeze-dried protective agent contains 130 g/L skimmed milk powder.
  • the culture medium contains water which accounts for 87.7% of the total mass of the medium, skim milk which accounts for 10% of the total mass of the medium, glucose which accounts for 0.5% of the total mass of the medium, and the total mass of the medium. 1.5% tryptone and 0.3% yeast extract of the total mass of the culture medium.
  • the pH of the culture medium is 6.8.
  • the present invention also provides a product for preventing and/or treating Helicobacter pylori infection, the product contains the Lactobacillus rhamnosus CCFM1119.
  • the number of viable cells of the Lactobacillus rhamnosus CCFM1119 is not less than 5 ⁇ 10 9 CFU/mL or 5 ⁇ 10 9 CFU/g.
  • the product includes food or medicine.
  • the medicine contains the aforementioned Lactobacillus rhamnosus CCFM1119, a drug carrier and/or a pharmaceutical excipient.
  • the drug carrier comprises microcapsules, microspheres, nanoparticles and/or liposomes.
  • the pharmaceutical excipients include excipients and/or additives.
  • the excipients include binders, fillers, disintegrants and/or lubricants.
  • the additive includes a solubilizer, a co-solvent, a latent solvent, and/or a preservative.
  • the dosage form of the medicine is powder, granule, capsule, tablet, pill or oral liquid.
  • the food is a health food; or the food is a dairy product, a bean product, or a fruit and vegetable product produced using a starter containing the above-mentioned Lactobacillus rhamnosus CCFM1119; or the food is Beverages or snacks containing the above-mentioned Lactobacillus rhamnosus CCFM1119.
  • the preparation method of the starter is to inoculate the above-mentioned Lactobacillus rhamnosus CCFM1119 into the culture medium at an inoculum of 2 to 4% of the total mass of the culture medium, and cultivate it at 37°C for 18 hours , Obtain the culture solution; centrifuge the culture solution to obtain the bacteria; wash the bacteria with physiological saline for 3 times and then resuspend with the freeze-dried protective agent to obtain the resuspension; the resuspension is lyophilized by the vacuum freezing method to obtain Starter.
  • the mass ratio of the freeze-dried protective agent to the bacterial cells is 2:1.
  • the freeze-dried protective agent contains 130 g/L skimmed milk powder.
  • the culture medium contains water which accounts for 87.7% of the total mass of the medium, skim milk which accounts for 10% of the total mass of the medium, glucose which accounts for 0.5% of the total mass of the medium, and the total mass of the medium. 1.5% tryptone and 0.3% yeast extract of the total mass of the culture medium.
  • the pH of the culture medium is 6.8.
  • the present invention provides a strain of Lactobacillus rhamnosus CCFM1119.
  • This Lactobacillus rhamnosus CCFM1119 can inhibit Helicobacter pylori, which is specifically embodied in:
  • the size of the inhibition zone of the supernatant of Lactobacillus rhamnosus CCFM1119 against Helicobacter pylori can reach 12.92mm;
  • Lactobacillus rhamnosus CCFM1119 can significantly reduce the adhesion of Helicobacter pylori to AGS cells
  • Lactobacillus rhamnosus CCFM1119 has great application prospects in inhibiting Helicobacter pylori (not for the purpose of disease diagnosis and treatment) and preparing Helicobacter pylori inhibitors.
  • the present invention provides a strain of Lactobacillus rhamnosus CCFM1119.
  • This Lactobacillus rhamnosus CCFM1119 has the effect of preventing and/or treating Helicobacter pylori infection, which is specifically embodied in:
  • Lactobacillus rhamnosus CCFM1119 can significantly alleviate the gastrointestinal symptoms of patients with Helicobacter pylori infection;
  • Lactobacillus rhamnosus CCFM1119 can significantly reduce the colonization of Helicobacter pylori in patients with Helicobacter pylori infection;
  • Lactobacillus rhamnosus CCFM1119 can significantly improve the clearance rate of Helicobacter pylori in patients with Helicobacter pylori infection.
  • Lactobacillus rhamnosus CCFM1119 has great application prospects in the preparation of products for preventing and/or treating Helicobacter pylori infection (such as food or medicine, etc.).
  • Lactobacillus rhamnosus (Lactobacillus rhamnosus) is a kind of probiotics. It has been included in the "List of Strains that Can be Used in Food" issued by the Ministry of Health. It can be seen that the active ingredient of the present invention is Lactobacillus rhamnosus CCFM1119 The product will not cause Helicobacter pylori to develop drug resistance, and at the same time, it will not cause adverse reactions in patients during the treatment process.
  • Lactobacillus rhamnosus (Lactobacillus rhamnosus) CCFM1119, taxonomically named Lactobacillus rhamnosus, has been deposited in the Guangdong Provincial Microbial Culture Collection on May 6, 2020, the deposit number is GDMCC No: 61013, and the deposit address is Guangzhou 5th Floor, Building 59, Yard 100, Xianlie Middle Road, City.
  • Figure 1 Adhesion rate of Helicobacter pylori in different groups to AGS cells.
  • Figure 2 Changes in GSRS scores of Helicobacter pylori-positive patients in different groups.
  • Figure 3 Changes in 14C expiratory values of Helicobacter pylori-positive patients in different groups.
  • Figure 4 The effect of storage time on the number of viable bacteria of Lactobacillus rhamnosus CCFM1119 powder.
  • the Helicobacter pylori involved in the following examples is Helicobacter pylori SS1 from the NTCC National Type Culture Collection; the Lactobacillus rhamnosus L.GG involved in the following examples is from the United States Type Culture Collection (ATCC), the deposit number is ATCC 53103; the F12 liquid medium and fetal calf serum involved in the following examples were purchased from Gibco, USA; the NaCl involved in the following examples was purchased from Sinopharm Group; the following examples involved The phenol red and urea were purchased from Macleans; the Columbia medium involved in the following examples was purchased from OXOID, the United Kingdom; the sterile defiberized sheep blood involved in the following examples was purchased from Hangzhou Xinrui Company; the following examples The BHI liquid medium involved was purchased from Qingdao Haibo Company.
  • MRS solid medium g/L: peptone 10g/L, beef extract 10g/L, glucose 20g/L, sodium acetate 2g/L, yeast powder 5g/L, diammonium hydrogen citrate 2g/L, K 2 PO 4 ⁇ 3H 2 O 2.6g/L, MgSO 4 ⁇ 7H 2 O 0.1g/L, MnSO 4 0.05g/L, Tween 80 1mL/L, agar 20g/L.
  • MRS liquid medium g/L: peptone 10g/L, beef extract 10g/L, glucose 20g/L, sodium acetate 2g/L, yeast powder 5g/L, diammonium hydrogen citrate 2g/L, K 2 PO 4 ⁇ 3H 2 O 2.6g/L, MgSO 4 ⁇ 7H 2 O 0.1g/L, MnSO 4 0.05g/L, Tween 80 1mL/L.
  • the detection method of the number of viable bacteria adopt the national standard "GB 4789.35-2016 National Food Safety Standard Food Microbiology Detection Lactic Acid Bacteria Detection”.
  • Columbia blood agar medium 39g of Columbia medium solid powder is dissolved in 1L of water, sterilized at 121°C for 15 minutes, and after cooling to 55°C to 60°C, 7.5% (v/v) sterile defiberized sheep blood is added. Pour the plate after mixing.
  • Example 1 Screening and identification of Lactobacillus rhamnosus
  • the liquid culture medium was enriched to obtain the strain CCFM1119 (the original number of the strain was JS-SZ-2-1), and the strain was stored in a 30% glycerol tube.
  • the genome of CCFM1119 was extracted, and the 16S rDNA of CCFM1119 was amplified and sequenced (completed by Shanghai Shenggong Biological Engineering Co., Ltd.). After sequencing analysis, the 16S rDNA sequence of this strain is shown in SEQ ID NO.1. Comparison in GenBank showed that the strain was Lactobacillus rhamnosus, named Lactobacillus rhamnosus CCFM1119.
  • Example 2 The inhibitory effect of Lactobacillus rhamnosus on the growth of Helicobacter pylori
  • MRS liquid medium as a negative control, streak Lactobacillus rhamnosus CCFM1119 on MRS solid medium and culture for 48 hours at 37°C to obtain a single colony; pick a single colony and inoculate it in MRS liquid medium at 37°C Cultivate for 18 hours under the conditions for activation, and activate for two consecutive generations to obtain the activation solution; inoculate the activation solution in the MRS liquid medium at an inoculum of 2% (v/v), and incubate at 37°C for 18 hours to obtain the bacterial solution; After centrifugation at 8000g for 10 minutes, the bacterial solution was filtered with a 0.22 ⁇ m sterile filter membrane to obtain the supernatant; the supernatant was determined by the Oxford cup method, the size of the inhibition zone of Lactobacillus rhamnosus CCFM1119 against Helicobacter pylori , To indicate the effect of inhibiting the growth of Helicobacter pylori, the results of the determination
  • MRS liquid medium has no inhibition zone against Helicobacter pylori, while the inhibition zone of Lactobacillus rhamnosus CCFM1119 supernatant against Helicobacter pylori can reach 12.92mm, indicating that Lactobacillus rhamnosus CCFM1119 has Inhibit the growth of Helicobacter pylori.
  • Table 1 The size of the inhibition zone of Lactobacillus rhamnosus CCFM1119 against Helicobacter pylori
  • Example 3 The effect of Lactobacillus rhamnosus on the adhesion of Helicobacter pylori
  • the Helicobacter pylori cells were resuspended in F12 medium to a concentration of 1 ⁇ 10 7 CFU/mL to obtain Helicobacter pylori resuspension; Lactobacillus rhamnosus L.GG cells were resuspended to concentration in F12 medium 1 ⁇ 10 7 CFU/mL to obtain a resuspension of Lactobacillus rhamnosus L.GG; resuspend the cells of Lactobacillus rhamnosus CCFM1119 in F12 medium to a concentration of 1 ⁇ 10 7 CFU/mL to obtain a rat Lactobacillus plumose CCFM1119 resuspension.
  • the AGS cells were resuspended in F12 medium containing 5% (v/v) fetal bovine serum and then added to a 96-well plate (2 ⁇ 10 4 cells/well), at 37°C and 5% CO 2 Cultivate for 12-16h. After the AGS cells are in an adherent state, wash the AGS cells with PBS 3 times to remove dead cells; add the Helicobacter pylori resuspension to the washed AGS cells at 37°C, 5% CO 2 After culturing in an incubator for 2 hours, wash with PBS solution 3 times to remove unadsorbed Helicobacter pylori, and obtain Helicobacter pylori infected AGS cells;
  • Hp group The Helicobacter pylori infection AGS cells not treated with Lactobacillus rhamnosus L.GG and Lactobacillus rhamnosus CCFM1119 were used as the model group (Hp group);
  • the Helicobacter pylori-infected AGS cells treated with Lactobacillus rhamnosus L.GG and the Helicobacter pylori-infected AGS cells treated with Lactobacillus rhamnosus CCFM1119 were the experimental groups, named Hp+LGG group and Hp+CCFM1119 group, respectively ;
  • urease reagent 9g/L NaCl, 14 ⁇ g/mL phenol red, 20mM urea, pH 6.8
  • urease reagent 9g/L NaCl, 14 ⁇ g/mL phenol red, 20mM urea, pH 6.8
  • the adhesion rate measured by subtracting the absorbance of the blank group from the absorbance of the model group is 100%.
  • the difference between the absorbance of the remaining groups and the absorbance of the blank group is equal to
  • the relative adhesion rate is obtained by comparing the difference between the absorbance of the model group and the absorbance of the blank group after subtracting the absorbance of the blank group. The measurement results are shown in Figure 1.
  • Example 4 Effect of Lactobacillus rhamnosus on gastrointestinal symptoms of Helicobacter pylori positive patients
  • Lactobacillus rhamnosus CCFM1119 was streaked on MRS solid medium and cultured at 37°C for 48 hours to obtain a single colony; a single colony was picked and inoculated in MRS liquid medium, and cultured at 37°C for 18 hours for activation.
  • Two consecutive activations Inoculate the activation solution into the culture medium at an inoculum of 2% (v/v), and incubate at 37°C for 18 hours to obtain the bacterial solution; centrifuge the bacterial solution at 8000g for 10 minutes to obtain the bacterial sludge ; Wash the bacterial mud 3 times with saline and resuspend it with a protective agent to a concentration of 1 ⁇ 10 10 CFU/mL to obtain a bacterial suspension; pre-incubate the bacterial suspension at a temperature of 37°C for 60 minutes and then freeze-dry to obtain a mouse Lactobacillus plumose CCFM1119 bacterial powder;
  • the preparation method of the culture medium is as follows: use 87.7% of the total weight of the culture medium to dissolve 10% enzymatically hydrolyzed skim milk, 0.5% glucose, 1.5% tryptone and 0.3% yeast extract, and then adjust its pH to 6.8 , Get the culture medium;
  • the ingredients of the protective agent include: 130g/L skimmed milk powder.
  • the placebo group (Placebo) took placebo twice a day, and the Lactobacillus rhamnosus CCFM1119 group took bacterial powder twice a day. The entire experimental period was 1 month (the placebo and Lactobacillus rhamnosus bacterial powder were different in composition, The appearance and packaging of the product are the same, there is no obvious difference).
  • the two groups of patients filled out the Gastrointestinal Symptom Rating Scale (GSRS) before and after the experiment (see Table 3 for details on the Gastrointestinal Symptom Rating Scale). The average value of each group was calculated to characterize the gastrointestinal health status of each group. Evaluation The improvement of the patient’s gastrointestinal symptoms before and after the experiment is shown in Figure 2.
  • Example 5 Effect of Lactobacillus rhamnosus on Helicobacter pylori colonization and clearance rate in patients with Helicobacter pylori positive
  • Lactobacillus rhamnosus CCFM1119 was streaked on MRS solid medium and cultured at 37°C for 48 hours to obtain a single colony; a single colony was picked and inoculated in MRS liquid medium, and cultured at 37°C for 18 hours for activation.
  • Two consecutive activations Inoculate the activation solution into the culture medium at an inoculum of 2% (v/v), and incubate at 37°C for 18 hours to obtain the bacterial solution; centrifuge the bacterial solution at 8000g for 10 minutes to obtain the bacterial sludge ; Wash the bacterial mud 3 times with saline and resuspend it with a protective agent to a concentration of 1 ⁇ 10 10 CFU/mL to obtain a bacterial suspension; pre-incubate the bacterial suspension at a temperature of 37°C for 60 minutes and then freeze-dry to obtain a mouse Lactobacillus plumose CCFM1119 bacterial powder;
  • the preparation method of the culture medium is as follows: use 87.7% of the total weight of the culture medium to dissolve 10% enzymatically hydrolyzed skim milk, 0.5% glucose, 1.5% tryptone and 0.3% yeast extract, and then adjust its pH to 6.8 , Get the culture medium;
  • the ingredients of the protective agent include: 130g/L skimmed milk powder.
  • the placebo group (Placebo) took placebo twice a day, and the Lactobacillus rhamnosus CCFM1119 group took bacterial powder twice a day.
  • the entire experimental period was 1 month (the placebo and Lactobacillus rhamnosus bacterial powder were different in composition, The appearance and packaging of the product are the same, there is no obvious difference).
  • the 14C breath value of the placebo group and the Lactobacillus rhamnosus CCFM1119 group with positive infection of Helicobacter pylori were measured by the 14C breath reagent bag and the tester before and after the experiment, and the amount of Helicobacter pylori colonization in the patients was evaluated. And clearance rate, the measurement results are shown in Figure 3 and Table 4;
  • the judging standard for the amount of Helicobacter pylori colonization is: the decrease of the 14C expiratory value of the Helicobacter pylori-positive infection after the experiment compared with the 14C expiratory value of the Helicobacter pylori-positive infection before the start of the experiment;
  • the evaluation criteria for the clearance rate of Helicobacter pylori are: the clinical 14C expiratory value is 100, that is, the 14C expiratory value is greater than or equal to 100 for Helicobacter pylori positive, and the 14C expiratory value is less than 100 for Helicobacter pylori negative. After the end, whether the positive infection of Helicobacter pylori turns to negative is judged whether the clearance rate of the positive infection of Helicobacter pylori increases.
  • Lactobacillus rhamnosus CCFM1119 can be used to prepare bacterial powder.
  • the specific preparation process of bacterial powder is as follows:
  • Lactobacillus rhamnosus CCFM1119 was streaked on MRS solid medium and cultured at 37°C for 48 hours to obtain a single colony; a single colony was picked and inoculated in MRS liquid medium, and cultured at 37°C for 18 hours for activation.
  • Two consecutive activations Inoculate the activation solution into the culture medium at an inoculum of 2% (v/v) and cultivate for 18 hours at 37°C to obtain the bacterial solution; centrifuge the bacterial solution at 8000g for 10 minutes to obtain the bacterial slurry; The bacterial sludge was washed 3 times with normal saline and resuspended with a protective agent to a concentration of 1 ⁇ 10 10 CFU/mL to obtain a bacterial suspension; the bacterial suspension was pre-incubated at 37°C for 60 minutes and then lyophilized to obtain rhamnose Lactobacillus CCFM1119 bacterial powder;
  • the preparation method of the culture medium is as follows: use 87.7% of the total weight of the culture medium to dissolve 10% enzymatically hydrolyzed skim milk, 0.5% glucose, 1.5% tryptone and 0.3% yeast extract, and then adjust its pH to 6.8 , Get the culture medium;
  • the ingredients of the protective agent include: 130g/L skimmed milk powder.
  • Lactobacillus rhamnosus CCFM1119 can be used to prepare capsule products.
  • the specific preparation process of capsule products is as follows
  • Lactobacillus rhamnosus CCFM1119 was streaked on MRS solid medium and cultured at 37°C for 48 hours to obtain a single colony; a single colony was picked and inoculated in MRS liquid medium, and cultured at 37°C for 18 hours for activation.
  • Two consecutive activations Inoculate the activation solution into the culture medium at an inoculum of 2% (v/v) and cultivate for 18 hours at 37°C to obtain the bacterial solution; centrifuge the bacterial solution at 8000g for 10 minutes to obtain the bacterial slurry; The bacterial mud was washed 3 times with saline and resuspended with a protective agent to a concentration of 1 ⁇ 10 10 CFU/mL to obtain a bacterial suspension; the bacterial suspension was added to a sodium alginate solution with a concentration of 30 g/L to a concentration of 2 ⁇ 10 9 CFU/mL, stir well to make the cells of Lactobacillus rhamnosus CCFM1119 evenly dispersed in the sodium alginate solution to obtain a mixed solution; squeeze the mixed solution to a calcium chloride solution with a concentration of 20g/L After the gel particles to be formed are statically solidified for 30 minutes, the gel particles are collected by filtration; the collected gel particles are freeze-dried for
  • the preparation method of the culture medium is as follows: dissolve 10% enzymatically hydrolyzed skim milk, 0.5% glucose, 1.5% tryptone and 0.3% yeast extract with 87.7% water based on the total weight of the culture medium, and then adjust the pH to 6.8 ,Get the culture medium.
  • Lactobacillus rhamnosus CCFM1119 can be used to prepare tablets.
  • the specific preparation process of the tablets is as follows:
  • Lactobacillus rhamnosus CCFM1119 was streaked on MRS solid medium and cultured at 37°C for 48 hours to obtain a single colony; a single colony was picked and inoculated in MRS liquid medium, and cultured at 37°C for 18 hours for activation.
  • Two consecutive activations Inoculate the activation solution into the culture medium at an inoculum of 2% (v/v) and cultivate for 18 hours at 37°C to obtain the bacterial solution; centrifuge the bacterial solution at 8000g for 10 minutes to obtain the bacterial slurry; The bacterial sludge was washed 3 times with normal saline and resuspended with a protective agent to a concentration of 1 ⁇ 10 10 CFU/mL to obtain a bacterial suspension; the bacterial suspension was pre-incubated at 37°C for 60 minutes and then lyophilized to obtain rhamnose Lactobacillus CCFM1119 bacterial powder;
  • the preparation method of the culture medium is as follows: use 87.7% of the total weight of the culture medium to dissolve 10% enzymatically hydrolyzed skim milk, 0.5% glucose, 1.5% tryptone and 0.3% yeast extract, and then adjust its pH to 6.8 , Get the culture medium;
  • the ingredients of the protective agent include: 130g/L skimmed milk powder.
  • Lactobacillus rhamnosus CCFM1119 can be used to prepare fermented milk.
  • the specific preparation process of fermented milk is as follows:
  • Lactobacillus rhamnosus CCFM1119 was streaked on MRS solid medium and cultured at 37°C for 48 hours to obtain a single colony; a single colony was picked and inoculated in MRS liquid medium, and cultured at 37°C for 18 hours for activation.
  • Two consecutive activations Inoculate the activation solution into the culture medium at an inoculum of 2% (v/v) and cultivate for 18 hours at 37°C to obtain the bacterial solution; centrifuge the bacterial solution at 8000g for 10 minutes to obtain the bacterial slurry; The bacterial sludge was washed 3 times with normal saline and resuspended with a protective agent to a concentration of 1 ⁇ 10 10 CFU/mL to obtain a bacterial suspension; the bacterial suspension was pre-incubated at 37°C for 60 minutes and then lyophilized to obtain rhamnose Lactobacillus CCFM1119 bacterial powder;
  • the preparation method of the culture medium is as follows: use 87.7% of the total weight of the culture medium to dissolve 10% enzymatically hydrolyzed skim milk, 0.5% glucose, 1.5% tryptone and 0.3% yeast extract, and then adjust its pH to 6.8 , Get the culture medium;
  • the ingredients of the protective agent include: 130g/L skimmed milk powder.
  • Lactobacillus rhamnosus CCFM1119 can be used to prepare soy milk.
  • the specific preparation process of soy milk is as follows:
  • Lactobacillus rhamnosus CCFM1119 was streaked on MRS solid medium and cultured at 37°C for 48 hours to obtain a single colony; a single colony was picked and inoculated in MRS liquid medium, and cultured at 37°C for 18 hours for activation.
  • Two consecutive activations Inoculate the activation solution into the culture medium at an inoculum of 2% (v/v) and cultivate for 18 hours at 37°C to obtain the bacterial solution; centrifuge the bacterial solution at 8000g for 10 minutes to obtain the bacterial slurry; The bacterial sludge was washed 3 times with normal saline and resuspended with a protective agent to a concentration of 1 ⁇ 10 10 CFU/mL to obtain a bacterial suspension; the bacterial suspension was pre-incubated at 37°C for 60 minutes and then lyophilized to obtain rhamnose Lactobacillus CCFM1119 bacterial powder;
  • the preparation method of the culture medium is as follows: use 87.7% of the total weight of the culture medium to dissolve 10% enzymatically hydrolyzed skim milk, 0.5% glucose, 1.5% tryptone and 0.3% yeast extract, and then adjust its pH to 6.8 , Get the culture medium;
  • the ingredients of the protective agent include: 130g/L skimmed milk powder.
  • Lactobacillus rhamnosus CCFM1119 can be used to prepare fruit and vegetable drinks.
  • the specific preparation process of fruit and vegetable drinks is as follows:
  • Lactobacillus rhamnosus CCFM1119 was streaked on MRS solid medium and cultured at 37°C for 48 hours to obtain a single colony; a single colony was picked and inoculated in MRS liquid medium, and cultured at 37°C for 18 hours for activation.
  • Two consecutive activations Inoculate the activation solution into the culture medium at an inoculum of 2% (v/v) and cultivate for 18 hours at 37°C to obtain the bacterial solution; centrifuge the bacterial solution at 8000g for 10 minutes to obtain the bacterial slurry; The bacterial sludge was washed 3 times with normal saline and resuspended with a protective agent to a concentration of 1 ⁇ 10 10 CFU/mL to obtain a bacterial suspension; the bacterial suspension was pre-incubated at 37°C for 60 minutes and then lyophilized to obtain rhamnose Lactobacillus CCFM1119 bacterial powder;
  • the preparation method of the culture medium is as follows: use 87.7% of the total weight of the culture medium to dissolve 10% enzymatically hydrolyzed skim milk, 0.5% glucose, 1.5% tryptone and 0.3% yeast extract, and then adjust its pH to 6.8 , Get the culture medium;
  • the ingredients of the protective agent include: 130g/L skimmed milk powder.
  • Lactobacillus rhamnosus CCFM1119 can be used to prepare milk drinks.
  • the specific preparation process of milk drinks is as follows:
  • Lactobacillus rhamnosus CCFM1119 was streaked on MRS solid medium and cultured at 37°C for 48 hours to obtain a single colony; a single colony was picked and inoculated in MRS liquid medium, and cultured at 37°C for 18 hours for activation.
  • Two consecutive activations Inoculate the activation solution into the culture medium at an inoculum of 2% (v/v) and cultivate for 18 hours at 37°C to obtain the bacterial solution; centrifuge the bacterial solution at 8000g for 10 minutes to obtain the bacterial slurry; The bacterial sludge was washed 3 times with normal saline and resuspended with a protective agent to a concentration of 1 ⁇ 10 10 CFU/mL to obtain a bacterial suspension; the bacterial suspension was pre-incubated at 37°C for 60 minutes and then lyophilized to obtain rhamnose Lactobacillus CCFM1119 bacterial powder;
  • the preparation method of the culture medium is as follows: use 87.7% of the total weight of the culture medium to dissolve 10% enzymatically hydrolyzed skim milk, 0.5% glucose, 1.5% tryptone and 0.3% yeast extract, and then adjust its pH to 6.8 , Get the culture medium;
  • the ingredients of the protective agent include: 130g/L skimmed milk powder.
  • the skimmed milk is heat sterilized at 95°C for 20 minutes and then cooled to 4°C to obtain the raw material; add Lactobacillus rhamnosus CCFM1119 powder to the raw material to a concentration of not less than 1 ⁇ 10 6 CFU/mL to obtain a milk beverage (milk beverage) It needs to be stored under refrigeration at 4°C).
  • Lactobacillus rhamnosus CCFM1119 can be used to prepare chocolate.
  • the specific preparation process of chocolate is as follows:
  • Lactobacillus rhamnosus CCFM1119 was streaked on MRS solid medium and cultured at 37°C for 48 hours to obtain a single colony; a single colony was picked and inoculated in MRS liquid medium, and cultured at 37°C for 18 hours for activation.
  • Two consecutive activations Inoculate the activation solution into the culture medium at an inoculum of 2% (v/v) and cultivate for 18 hours at 37°C to obtain the bacterial solution; centrifuge the bacterial solution at 8000g for 10 minutes to obtain the bacterial slurry; The bacterial sludge was washed 3 times with normal saline and resuspended with a protective agent to a concentration of 1 ⁇ 10 10 CFU/mL to obtain a bacterial suspension; the bacterial suspension was pre-incubated at 37°C for 60 minutes and then lyophilized to obtain rhamnose Lactobacillus CCFM1119 bacterial powder;
  • the preparation method of the culture medium is as follows: use 87.7% of the total weight of the culture medium to dissolve 10% enzymatically hydrolyzed skim milk, 0.5% glucose, 1.5% tryptone and 0.3% yeast extract, and then adjust its pH to 6.8 , Get the culture medium;
  • the ingredients of the protective agent include: 130g/L skimmed milk powder.
  • emulsifier liquid lecithin, soybean phospholipid, sorbitan monolaurate

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Mycology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Veterinary Medicine (AREA)
  • Polymers & Plastics (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Food Science & Technology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Nutrition Science (AREA)
  • Virology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Molecular Biology (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Epidemiology (AREA)
  • Pediatric Medicine (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

一株鼠李糖乳杆菌CCFM1119，其在抑制幽门螺杆菌中的应用及其在制备预防和/或治疗幽门螺杆菌感染的产品中的应用，该鼠李糖乳杆菌CCFM1119可显著缓解幽门螺杆菌感染患者的胃肠症状；降低幽门螺杆菌感染患者体内幽门螺杆菌定殖量；提高幽门螺杆菌感染患者体内幽门螺杆菌清除率。

Description

一株鼠李糖乳杆菌及其在抑制幽门螺杆菌中的应用 技术领域
本发明涉及一株鼠李糖乳杆菌及其在抑制幽门螺杆菌中的应用，属于微生物技术领域以及医药技术领域。
背景技术
幽门螺杆菌(Helicobacter pylori，Hp)是定殖在胃粘膜表面和十二指肠部位的一种微需氧革兰氏阴性菌。其最先被Marshall和Warren发现，两位学者为此获得了2005年诺贝尔生理或医学奖。幽门螺杆菌是最主要的流行性致病菌之一，其在全球人群中的检出率已经超过50％，经济发展水平低下和糟糕的卫生条件是螺杆菌感染流行的两个主要因素。在发达国家，成年人幽门螺杆菌的感染率约为30～40％，在发展中国家，成年人幽门螺杆菌的感染率高达80～90％左右。
幽门螺杆菌感染后一般难以自发清除而导致终身感染，除非进行根除治疗，或胃黏膜发生严重肠化生时细菌难以定殖，幽门螺杆菌才会自动消失。研究表明，幽门螺杆菌长期感染会引起慢性胃炎和十二指肠溃疡，并最终会发展成为胃癌。胃肠症状(Gastrointestinal Symptom Rating Scale，GSRS)是评估胃部健康症状的重要指标，其包含腹痛、酸反流、恶心和呕吐、腹鸣、嗳气、排便增多、排便不尽感等15种症状。临床研究表明，幽门螺杆菌感染往往伴随着餐后嗳气、恶心、腹胀、腹部不适等多种胃肠疾病症状，这些胃肠疾病症状大大影响了患者的生活质量。因此，恢复幽门螺杆菌感染导致的胃肠症状改变对改善患者的日常生活具有重要意义。
目前，主要是通过抗生素结合的三联或者四联疗法清除患者体内的幽门螺杆菌以恢复幽门螺杆菌感染导致的胃肠症状。但是，上述治疗方法由于频繁使用抗生素，易导致幽门螺杆菌耐药性增加，并且，使用上述治疗方法治疗患者的过程中，患者常常会出现严重的不良反应(如腹痛、恶心、腹泻等)，导致治疗有效率降低，治疗效果往往达不到预期。
因此，仍然需要一种药物或治疗方式，既不会增加幽门螺杆菌耐药性，同时，在治疗过程中不会导致患者产生不良反应，以提高幽门螺杆菌临床治疗的效果。
发明内容
[技术问题]
本发明要解决的技术问题是提供一株能够抑制幽门螺杆菌的鼠李糖乳杆菌(Lactobacillus rhamnosus)。
[技术方案]
为解决上述问题，本发明提供了一株鼠李糖乳杆菌(Lactobacillus rhamnosus)CCFM1119，所述鼠李糖乳杆菌CCFM1119保藏于广东省微生物菌种保藏中心，保藏编号为GDMCC No：61013，保藏日期为2020年05月06日。
所述鼠李糖乳杆菌CCFM1119来源于江苏省昆山市的健康人群新鲜粪便样本，该菌株经测序分析，其16S rDNA序列如SEQ ID NO.1所示，将测序得到的序列在GeneBank中进行核酸序列比对，结果显示菌株为鼠李糖乳杆菌，命名为鼠李糖乳杆菌CCFM1119。
所述鼠李糖乳杆菌CCFM1119在MRS固体培养基上的菌落呈乳白色半圆形凸起，表面光滑、湿润，边缘整齐。
本发明还提供了上述鼠李糖乳杆菌CCFM1119在抑制幽门螺杆菌方面不以疾病的诊断和治疗为目的的应用。
本发明还提供了一种幽门螺杆菌抑制剂，所述幽门螺杆菌抑制剂含有上述鼠李糖乳杆菌CCFM1119。
本发明还提供了上述鼠李糖乳杆菌CCFM1119在制备预防和/或治疗幽门螺杆菌感染的产品中的应用。
本发明的一种实施方式中，所述产品中，上述鼠李糖乳杆菌CCFM1119的活菌数为不低于5×10 9CFU/mL或5×10 9CFU/g。
本发明的一种实施方式中，所述产品包含食品或药品。
本发明的一种实施方式中，所述药品含有上述鼠李糖乳杆菌CCFM1119、药物载体和/或药用辅料。
本发明的一种实施方式中，所述药物载体包含微囊、微球、纳米粒和/或脂质体。
本发明的一种实施方式中，所述药用辅料包含赋形剂和/或附加剂。
本发明的一种实施方式中，所述赋形剂包含黏合剂、填充剂、崩解剂和/或润滑剂。
本发明的一种实施方式中，所述附加剂包含增溶剂、助溶剂、潜溶剂和/或防腐剂。
本发明的一种实施方式中，所述药品的剂型为粉剂、颗粒剂、胶囊剂、片剂、丸剂或口服液。
本发明的一种实施方式中，所述食品为保健食品；或所述食品为使用含有上述鼠李糖乳杆菌CCFM1119的发酵剂生产得到的乳制品、豆制品或果蔬制品；或所述食品为含有上述鼠李糖乳杆菌CCFM1119的饮料或零食。
本发明的一种实施方式中，所述发酵剂的制备方法为将上述鼠李糖乳杆菌CCFM1119按 照占培养基总质量2～4％的接种量接种到培养基中，于37℃下培养18h，得到培养液；将培养液离心，得到菌体；将菌体用生理盐水清洗3次后用冻干保护剂重悬，得到重悬液；将重悬液采用真空冷冻法进行冻干，得到发酵剂。
本发明的一种实施方式中，所述冻干保护剂和菌体的质量比为2:1。
在本发明的一种实施方式中，所述冻干保护剂包含130g/L的脱脂奶粉。
本发明的一种实施方式中，所述培养基包含占培养基总质量87.7％的水、占培养基总质量10％的脱脂乳、占培养基总质量0.5％的葡萄糖、占培养基总质量1.5％的胰蛋白胨以及占培养基总质量0.3％的酵母浸膏。
本发明的一种实施方式中，所述培养基的pH为6.8。
本发明还提供了一种用于预防和/或治疗幽门螺杆菌感染的产品，所述产品含有所述鼠李糖乳杆菌CCFM1119。
本发明的一种实施方式中，所述产品中，上述鼠李糖乳杆菌CCFM1119的活菌数为不低于5×10 9CFU/mL或5×10 9CFU/g。
本发明的一种实施方式中，所述产品包含食品或药品。
本发明的一种实施方式中，所述药品含有上述鼠李糖乳杆菌CCFM1119、药物载体和/或药用辅料。
本发明的一种实施方式中，所述药物载体包含微囊、微球、纳米粒和/或脂质体。
本发明的一种实施方式中，所述药用辅料包含赋形剂和/或附加剂。
本发明的一种实施方式中，所述赋形剂包含黏合剂、填充剂、崩解剂和/或润滑剂。
本发明的一种实施方式中，所述附加剂包含增溶剂、助溶剂、潜溶剂和/或防腐剂。
本发明的一种实施方式中，所述药品的剂型为粉剂、颗粒剂、胶囊剂、片剂、丸剂或口服液。
本发明的一种实施方式中，所述食品为保健食品；或所述食品为使用含有上述鼠李糖乳杆菌CCFM1119的发酵剂生产得到的乳制品、豆制品或果蔬制品；或所述食品为含有上述鼠李糖乳杆菌CCFM1119的饮料或零食。
本发明的一种实施方式中，所述发酵剂的制备方法为将上述鼠李糖乳杆菌CCFM1119按照占培养基总质量2～4％的接种量接种到培养基中，于37℃下培养18h，得到培养液；将培养液离心，得到菌体；将菌体用生理盐水清洗3次后用冻干保护剂重悬，得到重悬液；将重悬液采用真空冷冻法进行冻干，得到发酵剂。
本发明的一种实施方式中，所述冻干保护剂和菌体的质量比为2:1。
在本发明的一种实施方式中，所述冻干保护剂包含130g/L的脱脂奶粉。
本发明的一种实施方式中，所述培养基包含占培养基总质量87.7％的水、占培养基总质量10％的脱脂乳、占培养基总质量0.5％的葡萄糖、占培养基总质量1.5％的胰蛋白胨以及占培养基总质量0.3％的酵母浸膏。
本发明的一种实施方式中，所述培养基的pH为6.8。
有益效果：
1、本发明提供了一株鼠李糖乳杆菌(Lactobacillus rhamnosus)CCFM1119，此鼠李糖乳杆菌CCFM1119能够抑制幽门螺杆菌，具体体现在：
(1)鼠李糖乳杆菌CCFM1119上清液对幽门螺杆菌的抑菌圈大小可达12.92mm；
(2)鼠李糖乳杆菌CCFM1119可显著降低幽门螺杆菌对AGS细胞的粘附力，
可见，鼠李糖乳杆菌(Lactobacillus rhamnosus)CCFM1119在抑制幽门螺杆菌(不以疾病的诊断和治疗为目的)以及制备幽门螺杆菌抑制剂方面具有巨大的应用前景。
2、本发明提供了一株鼠李糖乳杆菌(Lactobacillus rhamnosus)CCFM1119，此鼠李糖乳杆菌CCFM1119具有预防和/或治疗幽门螺杆菌感染的作用，具体体现在：
(1)鼠李糖乳杆菌CCFM1119可显著缓解幽门螺杆菌感染患者的胃肠症状；
(2)鼠李糖乳杆菌CCFM1119可显著降低幽门螺杆菌感染患者体内幽门螺杆菌定殖量；
(3)鼠李糖乳杆菌CCFM1119可显著提高幽门螺杆菌感染患者体内幽门螺杆菌清除率，
可见，鼠李糖乳杆菌(Lactobacillus rhamnosus)CCFM1119在制备预防和/或治疗幽门螺杆菌感染的产品(如食品或药品等)中具有巨大的应用前景。
3、鼠李糖乳杆菌(Lactobacillus rhamnosus)是益生菌的一种，目前已被纳入卫生部下发的《可用于食品的菌种名单》，可见，本发明的有效成分为鼠李糖乳杆菌CCFM1119的产品不会使得幽门螺杆菌产生耐药性，同时，在治疗过程中不会导致患者产生不良反应。
生物材料保藏
一株鼠李糖乳杆菌(Lactobacillus rhamnosus)CCFM1119，分类学命名为Lactobacillus rhamnosus，已于2020年05月06日保藏于广东省微生物菌种保藏中心，保藏编号为GDMCC No：61013，保藏地址为广州市先烈中路100号大院59号楼5楼。
附图说明
图1：不同组别幽门螺杆菌粘附AGS细胞的粘附率。
图2：不同组别幽门螺杆菌阳性患者的GSRS评分变化情况。
图3：不同组别幽门螺杆菌阳性患者的14C呼气值变化情况。
图4：储存时间对鼠李糖乳杆菌CCFM1119菌粉活菌数的影响。
具体实施方式
下述实施例中涉及的幽门螺杆菌为来源于NTCC国家典型培养物保藏中心的幽门螺杆菌SS1；下述实施例中涉及的鼠李糖乳杆菌L.GG来源于美国模式培养物集存库(ATCC)，保藏编号为ATCC 53103；下述实施例中涉及的F12液体培养基和胎牛血清购自美国Gibco公司；下述实施例中涉及的NaCl购自国药集团；下述实施例中涉及的苯酚红与尿素购自麦克林公司；下述实施例中涉及的哥伦比亚培养基购自英国OXOID公司；下述实施例中涉及的无菌脱纤维绵羊血购自杭州新锐公司；下述实施例中涉及的BHI液体培养基购自青岛海博公司。
下述实施例中涉及的培养基如下：
MRS固体培养基(g/L)：蛋白胨10g/L、牛肉膏10g/L、葡萄糖20g/L、乙酸钠2g/L、酵母粉5g/L、柠檬酸氢二铵2g/L、K 2PO 4·3H 2O 2.6g/L、MgSO 4·7H 2O 0.1g/L、MnSO 4 0.05g/L、吐温80 1mL/L、琼脂20g/L。
MRS液体培养基(g/L)：蛋白胨10g/L、牛肉膏10g/L、葡萄糖20g/L、乙酸钠2g/L、酵母粉5g/L、柠檬酸氢二铵2g/L、K 2PO 4·3H 2O 2.6g/L、MgSO 4·7H 2O 0.1g/L、MnSO 4 0.05g/L、吐温80 1mL/L。
下述实施例中涉及的检测方法如下：
活菌数的检测方法：采用国标《GB 4789.35-2016食品安全国家标准食品微生物学检测乳酸菌检测》。
下述实施例中涉及的幽门螺杆菌菌体的制备方法如下：
将幽门螺杆菌在哥伦比亚血琼脂培养基上划线后，于37℃三气培养箱中(85％N 2、10％CO 2、5％O 2)培养3天，得到单菌落；挑取单菌落接种于含5％(v/v)胎牛血清的BHI培养基中，于37℃三气培养箱中(85％N 2、10％CO 2、5％O 2)培养4天得到种子液；将种子液以2％(v/v)的接种量接种于BHI液体培养基中，于37℃三气培养箱中(85％N 2、10％CO 2、5％O 2)培养4天，得到幽门螺杆菌菌液；将幽门螺杆菌菌液在8000g条件下离心10min，过滤，得到幽门螺杆菌菌体；
其中，哥伦比亚血琼脂培养基：39g哥伦比亚培养基固体粉末溶于1L水中，121℃灭菌15min，待冷却到55℃～60℃后加入7.5％(v/v)的无菌脱纤维绵羊血，混匀后倒板。
下述实施例中涉及的鼠李糖乳杆菌菌体的制备方法如下：
将鼠李糖乳杆菌划线于MRS固体培养基上，37℃条件下培养48h，得到单菌落；挑取单菌落接种于MRS液体培养基中，37℃条件下培养18h进行活化，连续活化两代，得到活化 液；将活化液按2％(v/v)的接种量接种于MRS液体培养基中，37℃条件下培养18h，得到菌液；将菌液在8000g条件下离心10min，过滤，得到鼠李糖乳杆菌菌体。
实施例1：鼠李糖乳杆菌的筛选及鉴定
1、筛选
以来源于江苏省昆山市的健康人群新鲜粪便为样本，将样本经预处理后，在30％左右甘油中保存于-80℃冰箱，取出解冻后，混匀样本，吸取0.5mL样本加到4.5mL 0.9％生理盐水进行梯度稀释，选择合适的梯度稀释液涂布在MRS固体培养基上，于37℃培养48h，挑取典型菌落至MRS平板上划线纯化，挑取单菌落转接至MRS液体培养基增菌，得到菌株CCFM1119(菌株原始编号为JS-SZ-2-1)，将菌株置于30％的甘油管中保藏。
2、鉴定
提取CCFM1119的基因组，将CCFM1119的16S rDNA进行扩增和测序(由上海生工生物工程股份有限公司完成)，经测序分析，该菌株的16S rDNA序列如SEQ ID NO.1所示，将该序列在GenBank中进行比对，结果显示菌株为鼠李糖乳杆菌，命名为鼠李糖乳杆菌(Lactobacillus rhamnosus)CCFM1119。
实施例2：鼠李糖乳杆菌对幽门螺杆菌生长的抑制作用
以MRS液体培养基为阴性对照，将鼠李糖乳杆菌CCFM1119划线于MRS固体培养基上，37℃条件下培养48h，得到单菌落；挑取单菌落接种于MRS液体培养基中，37℃条件下培养18h进行活化，连续活化两代，得到活化液；将活化液按2％(v/v)的接种量接种于MRS液体培养基中，37℃条件下培养18h，得到菌液；将菌液在8000g条件下离心10min后，采用0.22μm无菌滤膜进行过滤，得到上清液；用牛津杯法测定上清液中，鼠李糖乳杆菌CCFM1119对幽门螺杆菌的抑菌圈大小，以表示抑制幽门螺杆菌生长的效果，测定结果见表1(牛津杯法具体可参见文献：张婷婷，翟齐啸，金星等.具有拮抗空肠弯曲杆菌能力鸡源乳酸菌的筛选及特性.微生物学通报,2017,(44):118-125)。
由表1可知，MRS液体培养基对幽门螺杆菌无抑菌圈，而鼠李糖乳杆菌CCFM1119上清液对幽门螺杆菌的抑菌圈大小可达12.92mm，表明鼠李糖乳杆菌CCFM1119具有抑制幽门螺杆菌生长的作用。
表1鼠李糖乳杆菌CCFM1119对幽门螺杆菌的抑菌圈大小
组别 pH 抑菌圈大小(mm)
阴性对照 6.20 0
CCFM1119 3.54 12.92±0.19
实施例3：鼠李糖乳杆菌对幽门螺杆菌粘附力的影响
具体步骤如下：
(1)重悬液的制备
将幽门螺杆菌菌体用F12培养基重悬至浓度为1×10 7CFU/mL，得到幽门螺杆菌重悬液；将鼠李糖乳杆菌L.GG菌体用F12培养基重悬至浓度为1×10 7CFU/mL，得到鼠李糖乳杆菌L.GG重悬液；将鼠李糖乳杆菌CCFM1119菌体用F12培养基重悬至浓度为1×10 7CFU/mL，得到鼠李糖乳杆菌CCFM1119重悬液。
(2)幽门螺杆菌感染的AGS细胞的制备
将AGS细胞用含5％(v/v)胎牛血清的F12培养基重悬后添加至96孔板中(2×10 4个/孔)，于37℃、5％CO 2的条件下进行培养12～16h，待AGS细胞处于贴壁状态后，用PBS洗涤AGS细胞3次去除死细胞；将幽门螺杆菌重悬液添加至洗涤后的AGS细胞中，于37℃、5％CO 2的培养箱中培养2h后，用PBS液洗涤3次，除去未吸附的幽门螺杆菌，得到幽门螺杆菌感染AGS细胞；
(3)以不经鼠李糖乳杆菌L.GG和鼠李糖乳杆菌CCFM1119处理且未感染幽门螺杆菌的AGS细胞为空白组；
以不经鼠李糖乳杆菌L.GG和鼠李糖乳杆菌CCFM1119处理的幽门螺杆菌感染AGS细胞为模型组(Hp组)；
以经鼠李糖乳杆菌L.GG处理的幽门螺杆菌感染AGS细胞和经鼠李糖乳杆菌CCFM1119处理的幽门螺杆菌感染AGS细胞为实验组，分别命名为Hp+LGG组和Hp+CCFM1119组；
在幽门螺杆菌感染AGS细胞中分别加入0.2mL鼠李糖乳杆菌L.GG重悬液或鼠李糖乳杆菌CCFM1119重悬液，于37℃、5％CO 2的培养箱中培养2h，得到经鼠李糖乳杆菌L.GG处理的幽门螺杆菌感染AGS细胞和经鼠李糖乳杆菌CCFM1119处理的幽门螺杆菌感染AGS细胞；将经鼠李糖乳杆菌L.GG处理的幽门螺杆菌感染AGS细胞和经鼠李糖乳杆菌CCFM1119处理的幽门螺杆菌感染AGS细胞分别用PBS液洗涤5次后，在经鼠李糖乳杆菌L.GG处理的幽门螺杆菌感染AGS细胞和经鼠李糖乳杆菌CCFM1119处理的幽门螺杆菌感染AGS细胞中分别加入200μL尿素酶试剂(9g/L NaCl、14μg/mL苯酚红、20mM尿素，pH 6.8)，于37℃、5％CO 2的培养箱中培养2h，得到培养液；
通过酶标仪测定不同组的培养液的在波长550nm处的吸光度，以模型组吸光度减去空白组吸光度测得的粘附率为100％，其余组吸光度减去空白组吸光度后所得差值与模型组吸光度减去空白组吸光度后所得差值相比得到相对粘附率，测定结果见图1。
由图1可知，经鼠李糖乳杆菌CCFM1119处理后，幽门螺杆菌对AGS细胞的粘附率显 著下降，从模型组(Hp组)的100％下降到约70％左右，而鼠李糖乳杆菌L.GG并没有明显的降低幽门螺杆菌对AGS细胞粘附率的作用，幽门螺杆菌的粘附率几乎没有变化。此结果表明，鼠李糖乳杆菌CCFM1119可有效降低幽门螺杆菌对AGS细胞的粘附力。
实施例4：鼠李糖乳杆菌对幽门螺杆菌阳性患者胃肠症状的影响
鼠李糖乳杆菌CCFM1119划线于MRS固体培养基上，37℃条件下培养48h，得到单菌落；挑取单菌落接种于MRS液体培养基中，37℃条件下培养18h进行活化，连续活化两代，得到活化液；将活化液按2％(v/v)的接种量接种于培养基中，37℃条件下培养18h，得到菌液；将菌液在8000g条件下离心10min，得到菌泥；将菌泥用生理盐水清洗3次后用保护剂重悬至浓度为1×10 10CFU/mL，得到菌悬液；将菌悬液在温度37℃下预培养60min后冻干，得到鼠李糖乳杆菌CCFM1119菌粉；
其中，培养基的制备方法为：使用以培养基总重量计87.7％的水将10％酶水解脱脂乳、0.5％葡萄糖、1.5％胰蛋白胨与0.3％酵母浸膏溶解，然后调整其pH为6.8，得到培养基；
保护剂的成分包含：130g/L脱脂奶粉。
招募26名幽门螺杆菌阳性感染者(招募人群分布情况见表2，两组人群基线情况差异无统计学意义)，将26名幽门螺杆菌阳性感染者随机分为2组，其中，安慰剂组(Placebo)13人，鼠李糖乳杆菌CCFM1119组(CCFM1119)13人。
安慰剂组(Placebo)每天服用安慰剂两次，鼠李糖乳杆菌CCFM1119组每天服用两次菌粉，整个实验周期为1个月(安慰剂和鼠李糖乳杆菌菌粉除成分不同外，产品的外观和包装均相同，无明显差别)。两组患者分别于实验开始前和结束后填写胃肠症状评定量表(GSRS)(胃肠症状评定量表具体可见表3)，计算各组平均值以表征各组的胃肠健康状况，评价实验前后患者胃肠症状的改善情况，测定结果见图2。
由图2可知，实验前后，安慰剂组(Placebo)幽门螺杆菌阳性感染者的GSRS评分都在5以上，而鼠李糖乳杆菌CCFM1119组(CCFM1119)幽门螺杆菌阳性感染者实验开始前的GSRS评分约为6，实验结束后的GSRS评分下降到2左右。此结果表明，鼠李糖乳杆菌CCFM1119可显著缓解幽门螺杆菌感染患者胃肠症状。
表2幽门螺杆菌阳性感染者招募人群分布情况
组别 人数(N) 年龄 男/女 饮酒/不饮酒 吸烟/不吸烟
Placebo 13 48.15±3.70 2/11 1/12 0/13
CCFM1119 13 51.67±4.26 4/9 2/11 0/13
表3胃肠症状评定量表
Figure PCTCN2021095678-appb-000001
Figure PCTCN2021095678-appb-000002
Figure PCTCN2021095678-appb-000003
Figure PCTCN2021095678-appb-000004
实施例5：鼠李糖乳杆菌对幽门螺杆菌阳性患者体内幽门螺杆菌定殖量和清除率的影响
鼠李糖乳杆菌CCFM1119划线于MRS固体培养基上，37℃条件下培养48h，得到单菌落；挑取单菌落接种于MRS液体培养基中，37℃条件下培养18h进行活化，连续活化两代，得到活化液；将活化液按2％(v/v)的接种量接种于培养基中，37℃条件下培养18h，得到菌液；将菌液在8000g条件下离心10min，得到菌泥；将菌泥用生理盐水清洗3次后用保护剂重悬至浓度为1×10 10CFU/mL，得到菌悬液；将菌悬液在温度37℃下预培养60min后冻干，得到鼠李糖乳杆菌CCFM1119菌粉；
其中，培养基的制备方法为：使用以培养基总重量计87.7％的水将10％酶水解脱脂乳、0.5％葡萄糖、1.5％胰蛋白胨与0.3％酵母浸膏溶解，然后调整其pH为6.8，得到培养基；
保护剂的成分包含：130g/L脱脂奶粉。
招募26名幽门螺杆菌阳性感染者(招募人群分布情况见表2，两组人群基线情况差异无统计学意义)，将26名幽门螺杆菌阳性感染者随机分为2组，其中，安慰剂组(Placebo)13人，鼠李糖乳杆菌CCFM1119组(CCFM1119)13人。
安慰剂组(Placebo)每天服用安慰剂两次，鼠李糖乳杆菌CCFM1119组每天服用两次菌粉，整个实验周期为1个月(安慰剂和鼠李糖乳杆菌菌粉除成分不同外，产品的外观和包装均相同，无明显差别)。分别于实验开始前和结束后通过14C呼气试剂袋和检测仪测定安慰剂组和鼠李糖乳杆菌CCFM1119组幽门螺杆菌阳性感染者的14C呼气值，评估患者体内幽门螺杆菌定殖量和清除率，测定结果见图3和表4；
其中，幽门螺杆菌定殖量的评判标准为：实验结束后幽门螺杆菌阳性感染者的14C呼气值较实验开始前幽门螺杆菌阳性感染者的14C呼气值的下降量；
幽门螺杆菌清除率的评判标准为：临床14C呼气值的临界值为100，即14C呼气值大于等于100为幽门螺杆菌阳性，14C呼气值低于100为幽门螺杆菌阴性，以实验结束后幽门螺杆菌阳性感染者是否转变为阴性评判幽门螺杆菌阳性感染者的清除率是否升高。
由图3可知，实验结束后，安慰剂组(Placebo)幽门螺杆菌阳性感染者的14C呼气值与实验开始前相比几乎没有变化，而鼠李糖乳杆菌CCFM1119组(CCFM1119)幽门螺杆菌阳 性感染者的14C呼气值与实验开始前相比降低了100左右，具有显著性差异。此结果表明，鼠李糖乳杆菌CCFM1119可显著降低幽门螺杆菌感染患者体内幽门螺杆菌定殖量。
由表4可知，实验结束后，安慰剂组(Placebo)13个人中有2个人变为幽门螺杆菌阴性，阴性率为15.38％，鼠李糖乳杆菌CCFM1119组13个人中有8个人变为幽门螺杆菌阴性，阴性率高达61.54％，显著高于安慰剂组。此结果表明，鼠李糖乳杆菌CCFM1119可显著提高幽门螺杆菌感染患者体内幽门螺杆菌清除率。
表4不同组别幽门螺杆菌阳性患者的清除率
组别 人数(N) 阳性n 阴性n 阴性率(％)
Placebo 13 11 2 15.38
CCFM1119 13 5 8 61.54*
注：*表示与安慰剂组相比差异显著(p<0.05)。
实施例6：鼠李糖乳杆菌的应用
鼠李糖乳杆菌CCFM1119可用于制备菌粉，菌粉的具体制备过程如下：
鼠李糖乳杆菌CCFM1119划线于MRS固体培养基上，37℃条件下培养48h，得到单菌落；挑取单菌落接种于MRS液体培养基中，37℃条件下培养18h进行活化，连续活化两代，得到活化液；将活化液按2％(v/v)的接种量接种于培养基中，37℃条件下培养18h，得到菌液；将菌液经8000g离心10min，得到菌泥；将菌泥用生理盐水清洗3次后用保护剂重悬至浓度为1×10 10CFU/mL，得到菌悬液；将菌悬液在温度37℃下预培养60min后冻干，得到鼠李糖乳杆菌CCFM1119菌粉；
其中，培养基的制备方法为：使用以培养基总重量计87.7％的水将10％酶水解脱脂乳、0.5％葡萄糖、1.5％胰蛋白胨与0.3％酵母浸膏溶解，然后调整其pH为6.8，得到培养基；
保护剂的成分包含：130g/L脱脂奶粉。
准确称取2g鼠李糖乳杆菌CCFM1119菌粉溶解于10mL无菌生理盐水中，得到原始菌悬液；取0.5mL原始菌悬液加入4.5mL无菌生理盐水中，混匀，此时原始菌悬液稀释10倍，记为“n＝10”，从稀释后的菌悬液中取0.5mL加入4.5mL无菌生理盐水中，此时原始菌悬液稀释100倍，记为“n＝10 2”，以此类推将原始菌悬液稀释至1.0×10 8倍；取0.1mL稀释倍数为1.0×10 6(n＝10 6)、1.0×107(n＝10 7)和1.0×10 8(n＝10 8)的菌悬液，分别用MRS固体培养基倒平板，厌氧箱中37℃培养2d～3d，进行活菌计数，每周测定一次，持续一个月，测定鼠李糖乳杆菌CCFM1119菌粉的储存稳定性，测定结果见图4。
由图4可知，鼠李糖乳杆菌CCFM1119菌粉的初始活菌数高于10 10CFU/袋，符合产品规格；储存一个月的过程中，鼠李糖乳杆菌CCFM1119菌粉的活菌数与初始相比没有明显的下 降，活菌数一直高于10 10CFU/袋，表明鼠李糖乳杆菌CCFM1119菌粉在一个月的短期储存过程中性能比较稳定。
实施例7：鼠李糖乳杆菌的应用
鼠李糖乳杆菌CCFM1119可用于制备胶囊制品，胶囊制品的具体制备过程如下
鼠李糖乳杆菌CCFM1119划线于MRS固体培养基上，37℃条件下培养48h，得到单菌落；挑取单菌落接种于MRS液体培养基中，37℃条件下培养18h进行活化，连续活化两代，得到活化液；将活化液按2％(v/v)的接种量接种于培养基中，37℃条件下培养18h，得到菌液；将菌液经8000g离心10min，得到菌泥；将菌泥用生理盐水清洗3次后用保护剂重悬至浓度为1×10 10CFU/mL，得到菌悬液；菌悬液添加至浓度为30g/L的海藻酸钠溶液中至浓度为2×10 9CFU/mL后，充分搅拌，使得鼠李糖乳杆菌CCFM1119的细胞均匀地分散于海藻酸钠溶液中，得到混合液；将混合液挤压到浓度为20g/L的氯化钙溶液中形成胶粒；待形成的胶粒静止固化30min后，过滤收集胶粒；将收集得到的胶粒进行冷冻干燥48h，得到粉剂；将粉剂装入到药用胶囊中，得到胶囊制品；
其中，培养基的制备方法为：使用以培养基总重量计87.7％的水将10％酶水解脱脂乳、0.5％葡萄糖、1.5％胰蛋白胨与0.3％酵母浸膏溶解，然后调整其pH为6.8，得到培养基。
实施例8：鼠李糖乳杆菌的应用
鼠李糖乳杆菌CCFM1119可用于制备片剂，片剂的具体制备过程如下：
鼠李糖乳杆菌CCFM1119划线于MRS固体培养基上，37℃条件下培养48h，得到单菌落；挑取单菌落接种于MRS液体培养基中，37℃条件下培养18h进行活化，连续活化两代，得到活化液；将活化液按2％(v/v)的接种量接种于培养基中，37℃条件下培养18h，得到菌液；将菌液经8000g离心10min，得到菌泥；将菌泥用生理盐水清洗3次后用保护剂重悬至浓度为1×10 10CFU/mL，得到菌悬液；将菌悬液在温度37℃下预培养60min后冻干，得到鼠李糖乳杆菌CCFM1119菌粉；
其中，培养基的制备方法为：使用以培养基总重量计87.7％的水将10％酶水解脱脂乳、0.5％葡萄糖、1.5％胰蛋白胨与0.3％酵母浸膏溶解，然后调整其pH为6.8，得到培养基；
保护剂的成分包含：130g/L脱脂奶粉。
称取鼠李糖乳杆菌CCFM1119菌粉25.7重量份、淀粉55.0重量份、纤维素衍生物4.5重量份、羧甲基淀粉钠12.0重量份、滑石粉0.8重量份、蔗糖1.0重量份与水1.0重量份，得到原材料；将原材料混合，得到湿颗粒；将湿颗粒用中南制药机械厂的压片机进行压片后使用青州市益康中药机械有限公司的小型药物干燥机进行干燥，得到片剂。
实施例9：鼠李糖乳杆菌的应用
鼠李糖乳杆菌CCFM1119可用于制备发酵乳，发酵乳的具体制备过程如下：
鼠李糖乳杆菌CCFM1119划线于MRS固体培养基上，37℃条件下培养48h，得到单菌落；挑取单菌落接种于MRS液体培养基中，37℃条件下培养18h进行活化，连续活化两代，得到活化液；将活化液按2％(v/v)的接种量接种于培养基中，37℃条件下培养18h，得到菌液；将菌液经8000g离心10min，得到菌泥；将菌泥用生理盐水清洗3次后用保护剂重悬至浓度为1×10 10CFU/mL，得到菌悬液；将菌悬液在温度37℃下预培养60min后冻干，得到鼠李糖乳杆菌CCFM1119菌粉；
其中，培养基的制备方法为：使用以培养基总重量计87.7％的水将10％酶水解脱脂乳、0.5％葡萄糖、1.5％胰蛋白胨与0.3％酵母浸膏溶解，然后调整其pH为6.8，得到培养基；
保护剂的成分包含：130g/L脱脂奶粉。
将鼠李糖乳杆菌CCFM1119菌粉与商业干粉发酵剂保加利亚乳杆菌和商业干粉发酵剂嗜热链球菌按照质量比1:1:1的比例混合，得到发酵剂；将糖添加至鲜奶中至浓度为50g/L，得到混合液；将混合液在65℃、20MPa的条件下进行均质后在95℃下保温杀菌5min，得到发酵原料；将发酵原料降温至35℃后以0.03％(v/v)的接种量将发酵剂接种至发酵原料中，于35℃下保温发酵16h，得到发酵乳；将发酵乳于42℃下放置4h进行凝乳后，在4℃下冷藏24h进行后熟，得到发酵乳成品。
实施例10：鼠李糖乳杆菌的应用
鼠李糖乳杆菌CCFM1119可用于制备豆奶，豆奶的具体制备过程如下：
鼠李糖乳杆菌CCFM1119划线于MRS固体培养基上，37℃条件下培养48h，得到单菌落；挑取单菌落接种于MRS液体培养基中，37℃条件下培养18h进行活化，连续活化两代，得到活化液；将活化液按2％(v/v)的接种量接种于培养基中，37℃条件下培养18h，得到菌液；将菌液经8000g离心10min，得到菌泥；将菌泥用生理盐水清洗3次后用保护剂重悬至浓度为1×10 10CFU/mL，得到菌悬液；将菌悬液在温度37℃下预培养60min后冻干，得到鼠李糖乳杆菌CCFM1119菌粉；
其中，培养基的制备方法为：使用以培养基总重量计87.7％的水将10％酶水解脱脂乳、0.5％葡萄糖、1.5％胰蛋白胨与0.3％酵母浸膏溶解，然后调整其pH为6.8，得到培养基；
保护剂的成分包含：130g/L脱脂奶粉。
将大豆在温度80℃下浸泡2h后去除大豆皮，得到去皮大豆；将去皮大豆沥去浸泡水后加沸水磨浆，得到豆浆；将豆浆在高于80℃的温度条件下保温12min，得到熟豆浆；将熟豆 浆用150目筛网过滤后离心分离，得到粗豆奶；将粗豆奶加热到温度140～150℃后迅速导入真空冷却室进行抽真空，使得粗豆奶中的异味物质随着水蒸汽迅速排出，得到熟豆奶；将熟豆奶降温至约37℃后在熟豆奶中添加鼠李糖乳杆菌CCFM1119菌粉至浓度为不低于1×10 6CFU/mL，得到豆奶(豆奶需在4℃下冷藏保存)。
实施例11：鼠李糖乳杆菌的应用
鼠李糖乳杆菌CCFM1119可用于制备果蔬饮料，果蔬饮料的具体制备过程如下：
鼠李糖乳杆菌CCFM1119划线于MRS固体培养基上，37℃条件下培养48h，得到单菌落；挑取单菌落接种于MRS液体培养基中，37℃条件下培养18h进行活化，连续活化两代，得到活化液；将活化液按2％(v/v)的接种量接种于培养基中，37℃条件下培养18h，得到菌液；将菌液经8000g离心10min，得到菌泥；将菌泥用生理盐水清洗3次后用保护剂重悬至浓度为1×10 10CFU/mL，得到菌悬液；将菌悬液在温度37℃下预培养60min后冻干，得到鼠李糖乳杆菌CCFM1119菌粉；
其中，培养基的制备方法为：使用以培养基总重量计87.7％的水将10％酶水解脱脂乳、0.5％葡萄糖、1.5％胰蛋白胨与0.3％酵母浸膏溶解，然后调整其pH为6.8，得到培养基；
保护剂的成分包含：130g/L脱脂奶粉。
将新鲜水果和蔬菜洗净后榨汁，得到果蔬汁；将果蔬汁在温度140℃下高温热杀菌2秒，得到杀菌后的果蔬汁；将杀菌后的果蔬汁降温至约37℃后在杀菌后的果蔬汁中添加鼠李糖乳杆菌CCFM1119菌粉至浓度为不低于1×10 6CFU/mL，得到果蔬饮料(果蔬饮料需在4℃下冷藏保存)。
实施例12：鼠李糖乳杆菌的应用
鼠李糖乳杆菌CCFM1119可用于制备乳饮品，乳饮品的具体制备过程如下：
鼠李糖乳杆菌CCFM1119划线于MRS固体培养基上，37℃条件下培养48h，得到单菌落；挑取单菌落接种于MRS液体培养基中，37℃条件下培养18h进行活化，连续活化两代，得到活化液；将活化液按2％(v/v)的接种量接种于培养基中，37℃条件下培养18h，得到菌液；将菌液经8000g离心10min，得到菌泥；将菌泥用生理盐水清洗3次后用保护剂重悬至浓度为1×10 10CFU/mL，得到菌悬液；将菌悬液在温度37℃下预培养60min后冻干，得到鼠李糖乳杆菌CCFM1119菌粉；
其中，培养基的制备方法为：使用以培养基总重量计87.7％的水将10％酶水解脱脂乳、0.5％葡萄糖、1.5％胰蛋白胨与0.3％酵母浸膏溶解，然后调整其pH为6.8，得到培养基；
保护剂的成分包含：130g/L脱脂奶粉。
将脱脂奶在95℃热杀菌20min后冷却至4℃，得到原料；在原料中添加鼠李糖乳杆菌CCFM1119菌粉至浓度为不低于1×10 6CFU/mL，得到乳饮品(乳饮品需在4℃下冷藏保存)。
实施例13：鼠李糖乳杆菌的应用
鼠李糖乳杆菌CCFM1119可用于制备巧克力，巧克力的具体制备过程如下：
鼠李糖乳杆菌CCFM1119划线于MRS固体培养基上，37℃条件下培养48h，得到单菌落；挑取单菌落接种于MRS液体培养基中，37℃条件下培养18h进行活化，连续活化两代，得到活化液；将活化液按2％(v/v)的接种量接种于培养基中，37℃条件下培养18h，得到菌液；将菌液经8000g离心10min，得到菌泥；将菌泥用生理盐水清洗3次后用保护剂重悬至浓度为1×10 10CFU/mL，得到菌悬液；将菌悬液在温度37℃下预培养60min后冻干，得到鼠李糖乳杆菌CCFM1119菌粉；
其中，培养基的制备方法为：使用以培养基总重量计87.7％的水将10％酶水解脱脂乳、0.5％葡萄糖、1.5％胰蛋白胨与0.3％酵母浸膏溶解，然后调整其pH为6.8，得到培养基；
保护剂的成分包含：130g/L脱脂奶粉。
将可可块和白砂糖以1:1～1:3的质量比混合后加热，搅拌均匀，得到巧克力融浆；先将乳化剂(液体卵磷脂、大豆磷脂、山梨醇酐单月桂酸酯)与鼠李糖乳杆菌CCFM1119菌粉以乳化剂：菌粉＝80～90：10～20的质量比混合均匀，再进行精磨、除酸、除水、结晶、调温，最后选择合适的模型浇注成型，得到巧克力(巧克力需在4℃下冷藏保存)。
虽然本发明已以较佳实施例公开如上，但其并非用以限定本发明，任何熟悉此技术的人，在不脱离本发明的精神和范围内，都可做各种的改动与修饰，因此本发明的保护范围应该以权利要求书所界定的为准。

Claims (20)

  1. 一株鼠李糖乳杆菌(Lactobacillus rhamnosus)，其特征在于，所述鼠李糖乳杆菌保藏于广东省微生物菌种保藏中心，保藏编号为GDMCC No.61013，保藏日期为2020年05月06日。
  2. 一种幽门螺杆菌抑制剂，其特征在于，所述幽门螺杆菌抑制剂含有权利要求1所述鼠李糖乳杆菌，所述幽门螺杆菌为幽门螺杆菌(Helicobacter pylori)SS1。
  3. 如权利要求1所述的幽门螺杆菌抑制剂，其特征在于，所述抑制剂中，所述鼠李糖乳杆菌的活菌数为不低于5×10 9CFU/mL或5×10 9CFU/g。
  4. 一种用于预防和/或治疗幽门螺杆菌SS1感染的产品，其特征在于，所述产品含有权利要求1所述的鼠李糖乳杆菌。
  5. 如权利要求4所述的一种用于预防和/或治疗幽门螺杆菌SS1感染的产品，其特征在于，所述产品中，所述鼠李糖乳杆菌的活菌数为不低于5×10 9CFU/mL或5×10 9CFU/g。
  6. 如权利要求5所述的一种用于预防和/或治疗幽门螺杆菌SS1感染的产品，其特征在于，所述产品为食品或药品。
  7. 如权利要求6所述的一种用于预防和/或治疗幽门螺杆菌SS1感染的产品，其特征在于，所述药品含有鼠李糖乳杆菌、药物载体和/或药用辅料。
  8. 如权利要求7所述的一种用于预防和/或治疗幽门螺杆菌SS1感染的产品，其特征在于，所述药品的剂型为粉剂、颗粒剂、胶囊剂、片剂、丸剂或口服液。
  9. 如权利要求6所述的一种用于预防和/或治疗幽门螺杆菌SS1感染的产品，其特征在于，所述食品为保健食品。
  10. 如权利要求6所述的一种用于预防和/或治疗幽门螺杆菌SS1感染的产品，其特征在于，所述食品为使用含有鼠李糖乳杆菌的发酵剂生产得到的乳制品、豆制品或果蔬制品。
  11. 如权利要求6所述的一种用于预防和/或治疗幽门螺杆菌SS1感染的产品，其特征在于，所述食品为含有鼠李糖乳杆菌的饮料或零食。
  12. 如权利要求10所述的一种用于预防和/或治疗幽门螺杆菌SS1感染的产品，其特征在于，所述发酵剂的制备方法为将鼠李糖乳杆菌按照占培养基总质量2～4％的接种量接种到培养基中，于37℃下培养18h，得到培养液；将培养液离心，得到菌体；将菌体用生理盐水清洗3次后用冻干保护剂重悬，得到重悬液；将重悬液采用真空冷冻法进行冻干，得到发酵剂。
  13. 如权利要求12所述的一种用于预防和/或治疗幽门螺杆菌SS1感染的产品，其特征在于，所述冻干保护剂和菌体的质量比为2:1。
  14. 如权利要求13所述的一种用于预防和/或治疗幽门螺杆菌SS1感染的产品，其特征在于，所述培养基为：占培养基总质量87.7％的水、占培养基总质量10％的脱脂乳、占培养基 总质量0.5％的葡萄糖、占培养基总质量1.5％的胰蛋白胨以及占培养基总质量0.3％的酵母浸膏。
  15. 如权利要求14所述的一种用于预防和/或治疗幽门螺杆菌SS1感染的产品，其特征在于，所述培养基的pH为6.8。
  16. 权利要求1所述鼠李糖乳杆菌在制备预防和/或治疗幽门螺杆菌SS1感染的产品中的应用。
  17. 如权利要求16所述的应用，其特征在于，所述产品中，所述鼠李糖乳杆菌的活菌数为不低于5×10 9CFU/mL或5×10 9CFU/g。
  18. 如权利要求17所述的应用，其特征在于，所述产品为食品或药品。
  19. 如权利要求18所述的应用，其特征在于，所述药品含有鼠李糖乳杆菌、药物载体和/或药用辅料。
  20. 如权利要求18所述的应用，其特征在于，所述食品为使用含有鼠李糖乳杆菌的发酵剂生产得到的乳制品、豆制品或果蔬制品。
PCT/CN2021/095678 2020-05-29 2021-05-25 一株鼠李糖乳杆菌及其在抑制幽门螺杆菌中的应用 WO2021238890A1 (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US17/711,257 US20220228106A1 (en) 2020-05-29 2022-04-01 Strain of Lactobacillus rhamnosus and Application thereof in Inhibiting Helicobacter pylori

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202010478076.0 2020-05-29
CN202010478076.0A CN111607538B (zh) 2020-05-29 2020-05-29 一株鼠李糖乳杆菌及其在抑制幽门螺杆菌中的应用

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US17/711,257 Continuation US20220228106A1 (en) 2020-05-29 2022-04-01 Strain of Lactobacillus rhamnosus and Application thereof in Inhibiting Helicobacter pylori

Publications (1)

Publication Number Publication Date
WO2021238890A1 true WO2021238890A1 (zh) 2021-12-02

Family

ID=72194164

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2021/095678 WO2021238890A1 (zh) 2020-05-29 2021-05-25 一株鼠李糖乳杆菌及其在抑制幽门螺杆菌中的应用

Country Status (3)

Country Link
US (1) US20220228106A1 (zh)
CN (1) CN111607538B (zh)
WO (1) WO2021238890A1 (zh)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115029265A (zh) * 2022-06-20 2022-09-09 自然资源部第一海洋研究所 一株植物乳杆菌m503及其与鼠李糖乳杆菌复配的制剂和在抗幽门螺杆菌药物中的应用
CN115226771A (zh) * 2022-07-19 2022-10-25 广州喜乐食品企业有限公司 一种抑制幽门螺旋杆菌的风味酸奶及其制备方法

Families Citing this family (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111607538B (zh) * 2020-05-29 2021-03-02 江南大学 一株鼠李糖乳杆菌及其在抑制幽门螺杆菌中的应用
CN112746034B (zh) * 2020-09-14 2022-07-29 河北一然生物科技股份有限公司 一种协同抑制幽门螺旋杆菌的鼠李糖乳杆菌lr863和鼠李糖乳杆菌lr519及其应用
CN112940984B (zh) * 2021-03-31 2023-07-25 江苏蓝泽生物科技有限公司 抗幽门螺杆菌、降血糖、调理肠胃和增加免疫力的复合乳酸杆菌制剂及其制备方法
CN114214230B (zh) * 2021-12-09 2023-08-08 江南大学 一株具有共聚幽门螺杆菌能力的北酸乳杆菌及其应用
CN115153026B (zh) * 2022-04-13 2024-01-26 微康益生菌(苏州)股份有限公司 鼠李糖乳杆菌菌株LRa05在制备抑制幽门螺杆菌和/或变异链球菌制品方面的用途
CN114958658B (zh) * 2022-05-11 2023-08-11 广西爱生生命科技有限公司 一株抑制幽门螺旋杆菌的鼠李糖乳杆菌a21149及其应用
CN116396884A (zh) * 2022-08-15 2023-07-07 江苏华熙益能生物科技有限公司 鼠李糖乳杆菌和一种抑制幽门螺杆菌的组合物
CN115851551B (zh) * 2022-12-29 2023-08-01 广西爱生生命科技有限公司 一株具有抗幽门螺杆菌作用的戊糖片球菌a21358及其应用
CN116083325B (zh) * 2023-03-22 2023-12-05 微康益生菌(苏州)股份有限公司 一种改善幽门螺杆菌相关性胃肠疾病的鼠李糖乳杆菌及其应用
CN116121155B (zh) * 2023-04-14 2023-09-05 微康益生菌(苏州)股份有限公司 一株鼠李糖乳酪杆菌LRa09及其应用与产品
CN116333945B (zh) * 2023-05-12 2023-08-01 成都海若健康科技有限公司 一种抗幽门螺杆菌的鼠李糖乳酪杆菌puk09及其应用、产品及方法
CN116640704B (zh) * 2023-07-21 2023-11-24 青岛蔚蓝生物集团有限公司 一株抑制幽门螺旋杆菌的戊糖片球菌及其应用

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100646938B1 (ko) * 2005-09-23 2006-11-23 일동제약주식회사 헬리코박터 필로리의 정착과 생육 활성을 저해하는락토바실루스 람노서스 아이디씨씨 3201
CN1982437A (zh) * 2005-12-16 2007-06-20 统一企业(中国)投资有限公司 一种新颖的耐酸耐胆盐、抗肠道致病菌和抗氧化能力鼠李糖乳杆菌株
CN101314763A (zh) * 2007-06-01 2008-12-03 统一企业(中国)投资有限公司 具有抗胃肠道致病菌、抗氧化和降血压作用的短双歧杆菌及其用途
CN110628660A (zh) * 2018-06-22 2019-12-31 景岳生物科技(中国)有限公司 一种乳酸菌组合物用于抑制胃幽门螺旋杆菌引发的胃炎及其用途
US20200077692A1 (en) * 2018-09-12 2020-03-12 Genmont Biotech Incorporation Lactobacillus composition for inhibiting gastritis induced by gastric helicobacter pylori and use thereof
CN111607538A (zh) * 2020-05-29 2020-09-01 江南大学 一株鼠李糖乳杆菌及其在抑制幽门螺杆菌中的应用

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI316545B (en) * 2006-08-09 2009-11-01 Uni President Entpr Corp Novel lactobacillus rhamnosus strain lv108 and uses thereof

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100646938B1 (ko) * 2005-09-23 2006-11-23 일동제약주식회사 헬리코박터 필로리의 정착과 생육 활성을 저해하는락토바실루스 람노서스 아이디씨씨 3201
CN1982437A (zh) * 2005-12-16 2007-06-20 统一企业(中国)投资有限公司 一种新颖的耐酸耐胆盐、抗肠道致病菌和抗氧化能力鼠李糖乳杆菌株
CN101314763A (zh) * 2007-06-01 2008-12-03 统一企业(中国)投资有限公司 具有抗胃肠道致病菌、抗氧化和降血压作用的短双歧杆菌及其用途
CN110628660A (zh) * 2018-06-22 2019-12-31 景岳生物科技(中国)有限公司 一种乳酸菌组合物用于抑制胃幽门螺旋杆菌引发的胃炎及其用途
US20200077692A1 (en) * 2018-09-12 2020-03-12 Genmont Biotech Incorporation Lactobacillus composition for inhibiting gastritis induced by gastric helicobacter pylori and use thereof
CN111607538A (zh) * 2020-05-29 2020-09-01 江南大学 一株鼠李糖乳杆菌及其在抑制幽门螺杆菌中的应用

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
ZUO FANGLEI, APPASWAMY AMULYA, GEBREMARIAM HANNA G., JONSSON ANN-BETH: "Role of Sortase A in Lactobacillus gasseri Kx110A1 Adhesion to Gastric Epithelial Cells and Competitive Exclusion of Helicobacter pylori", FRONTIERS IN MICROBIOLOGY, vol. 10, 1 January 2019 (2019-01-01), pages 2770, XP055871237, DOI: 10.3389/fmicb.2019.02770 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115029265A (zh) * 2022-06-20 2022-09-09 自然资源部第一海洋研究所 一株植物乳杆菌m503及其与鼠李糖乳杆菌复配的制剂和在抗幽门螺杆菌药物中的应用
CN115226771A (zh) * 2022-07-19 2022-10-25 广州喜乐食品企业有限公司 一种抑制幽门螺旋杆菌的风味酸奶及其制备方法
CN115226771B (zh) * 2022-07-19 2023-06-30 广州喜乐食品企业有限公司 一种抑制幽门螺旋杆菌的风味酸奶及其制备方法

Also Published As

Publication number Publication date
CN111607538B (zh) 2021-03-02
CN111607538A (zh) 2020-09-01
US20220228106A1 (en) 2022-07-21

Similar Documents

Publication Publication Date Title
WO2021238890A1 (zh) 一株鼠李糖乳杆菌及其在抑制幽门螺杆菌中的应用
WO2021238907A1 (zh) 一株能够预防和/或治疗幽门螺杆菌感染的卷曲乳杆菌
CN106834187B (zh) 一种两歧双歧杆菌及其用途
EP0508701B1 (en) Probiotic
US20100166721A1 (en) Probotic compositions and uses thereof
US20110165127A1 (en) Dairy-derived probiotic compositions and uses thereof
CN112322528B (zh) 一株可干预代谢综合征的鼠李糖乳杆菌及应用
CN112322527A (zh) 一株可干预代谢综合征的罗伊氏乳杆菌及应用
WO2005092122A1 (ja) ユッカ抽出物、キラヤ抽出物及び乳酸菌からなる組成物及び同組成物を含有する飲食品
CN113403231B (zh) 一株可干预代谢综合征的罗伊氏乳杆菌ccfm1178及应用
CN116024130B (zh) 一株降血尿酸发酵乳杆菌a21215及其应用
CN113943681B (zh) 一株降低炎症反应且具有缓解便秘作用的长双歧杆菌
CN110106103B (zh) 假小链双歧杆菌ccfm1047、其组合物、发酵食品、用途、菌剂及其菌剂制备方法
CN111471626A (zh) 一株能够抑制幽门螺杆菌的瑞士乳杆菌及其应用
CN110093287A (zh) 假小链双歧杆菌ccfm1045、其组合物、发酵食品、用途、菌剂及其菌剂制备方法
CN114540243A (zh) 一种能够预防和或治疗幽门螺杆菌感染的鼠李糖乳杆菌及其应用
CN114214230B (zh) 一株具有共聚幽门螺杆菌能力的北酸乳杆菌及其应用
CN114317334A (zh) 一株能够与幽门螺杆菌共聚集的清酒乳杆菌及其应用
WO2019174002A1 (zh) 戊糖片球菌ccfm1012、其发酵食品及其在制备拮抗空肠弯曲杆菌感染药物中的应用
CN116024129A (zh) 一株能够与幽门螺杆菌共聚集的卷曲乳杆菌及其应用
CN110106104B (zh) 假小链双歧杆菌ccfm1048、其组合物、发酵食品、用途、菌剂及其菌剂制备方法
CN114231449B (zh) 一株具有共聚幽门螺杆菌能力的嗜酸乳杆菌及其应用
CN114561325B (zh) 一株能够改变模拟胃肠道环境中胆汁酸含量且具有缓解便秘作用的长双歧杆菌及其应用
CN115721017B (zh) 一株可激活肠道抗炎靶点的短双歧杆菌及应用
CN115948296B (zh) 用于改善肌骨健康和/或提高运动能力的植物乳杆菌及其组合物与应用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21814136

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 21814136

Country of ref document: EP

Kind code of ref document: A1