Lactobacillus gasseri and application thereof

Abstract

The invention discloses lactobacillus gasseri WT002 which is preserved in the general microbiological center of the China Committee for Culture Collection of Microorganisms on February 18, 2025, and the preservation number of the lactobacillus gasseri WT002 is CGMCC No.33552. The lactobacillus gasseri WT002 is named as Lactobacillus gasseri WT002. The lactobacillus gasseri WT002 can inhibit the growth of various vaginal pathogenic bacteria, has strong vaginal epithelial cell and cervical cancer cell adhesion ability, can be used for preventing and / or relieving and / or treating genital tract diseases, and further can be used for preparing medicines, hygienic products and the like.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, and in particular to a type of Lactobacillus gasseri and its applications. Background Technology

[0002] Lactobacillus ( Lactobacillus Lactobacilli are widely present in the human body, especially in the female reproductive tract flora. They play a vital role in protecting vaginal health by producing substances such as lactic acid and hydrogen peroxide, which inhibit the growth of pathogenic bacteria. However, changes in external environmental factors can reduce the number of lactobacilli in the vagina, leading to an overgrowth of pathogenic bacteria and causing diseases such as vaginitis. Current treatments often involve the local application of antibiotics to inhibit bacterial growth, but with increasing antibiotic resistance, the recurrence rate of these diseases continues to rise. Summary of the Invention

[0003] This application provides a Lactobacillus gasseri that has a strong ability to produce acid and H2O2, and can inhibit the growth of various vaginal pathogens. It can be used to prevent and / or alleviate and / or treat diseases caused by vaginal flora imbalance.

[0004] This application provides a Lactobacillus gasseri ( Lactobacillus gasseri Lactobacillus gasseri WT002 was deposited on February 18, 2025, at the China General Microbiological Culture Collection Center (CGMCC, address: No. 3, No. 1, Beizhan West Road, Chaoyang District, Beijing), with accession number CGMCC No. 33552.

[0005] This application provides the use of the aforementioned Lactobacillus gasseri in the preparation of a medicament for the prevention and / or treatment of reproductive tract infections.

[0006] This application provides the use of the aforementioned Lactobacillus gasseri in the preparation of a drug for inhibiting pathogenic bacteria.

[0007] Furthermore, the pathogenic bacteria include at least one of Escherichia coli, Gardnerella vaginalis, Candida albicans, and Prevotella.

[0008] This application provides the use of the aforementioned Lactobacillus gasseri in the preparation of a drug, said drug having vaginal epithelial cell adhesion function.

[0009] This application provides the use of the aforementioned Lactobacillus gasseri in the preparation of a drug, said drug having the function of adhering to human cervical cancer cells.

[0010] This application provides the use of the aforementioned *Lactobacillus gasseri* in the preparation of a pharmaceutical product, the pharmaceutical product being used in a product for improving reproductive tract flora, wherein improving reproductive tract flora includes promoting the proliferation of beneficial vaginal bacteria, and promoting the proliferation of beneficial vaginal bacteria includes promoting *Lactobacillus curvaturei* (…). Lactobacillus crispatus ) proliferation, promoting the growth of inert lactobacilli ( Lactobacillus iners ) proliferation and promotion of Lactobacillus janniae ( Lactobacillus jensenii )proliferation.

[0011] This application provides a composition comprising the aforementioned Lactobacillus gasseri WT002 and optionally one or more pharmaceutically acceptable carriers.

[0012] This application provides an application of the aforementioned Lactobacillus gasseri or the aforementioned composition in the preparation of hygiene products.

[0013] The Lactobacillus gasseri disclosed in this application ( Lactobacillus gasseri Lactobacillus gasseri WT002, with accession number CGMCC No. 33552, exhibits strong growth viability and possesses advantages such as inhibiting the proliferation of vaginal pathogens, tolerance to low pH, high lactic acid production, and strong vaginal cell adhesion. It can significantly improve and regulate the vaginal microenvironment and inhibit vaginal pathogens. Therefore, Lactobacillus gasseri WT002 and / or its metabolites show promising application prospects in the preparation of products for the prevention and / or treatment of urogenital diseases (such as pharmaceuticals, medical devices, or hygiene products). Attached Figure Description

[0014] To more clearly illustrate the technical solutions of the embodiments of this application, the drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0015] Figure 1 The growth curve of Lactobacillus gasseri WT002 provided in the embodiments of this application. Detailed Implementation

[0016] The following specific embodiments illustrate the implementation of the invention. Those skilled in the art can easily understand other advantages and effects of the invention from the content disclosed in this specification.

[0017] Terminology Definition In this application, the term "Lactobacillus gasseri" (Lactobacillus gasseri) Lactobacillus gasseri ")" or "Lactobacillus gasseri ( L.crispatus "( )" usually refers to species in the genus *Lactobacillus*. This species is typically distinguished from other *Lactobacillus* species based on the polynucleotide sequence of the 16S rRNA gene of the ribosome.

[0018] In the present application, the term "sequence identity" generally refers to two polynucleotide or amino acid sequences that are the same (i.e., in nucleotides, or in residues) over the comparison window. The term "% sequence identity" is calculated as follows: over the comparison window, the number of positions at which the identical nucleic acid base (e.g., A, T, C, G, U, or I) or amino acid residue occurs in both sequences is ascertained, the number of matched positions is divided by the total number of positions in the comparison window (i.e., the comparison window size), and the result is multiplied by 100 to yield the percentage of sequence identity. The term "substantial identity" or "substantially the same" generally refers to a property of polynucleotide or amino acid sequences wherein the polynucleotide or amino acid sequences comprise at least about 85% sequence identity, such as at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, or 95% sequence identity, more usually at least about 96%, 97%, 98%, or 99% sequence identity, when compared to a reference sequence over a comparison window of at least 18 nucleotides (6 amino acids), specifically over a window of at least 18-48 nucleotides (6-16 amino acids), frequently over a window of at least 24-48 nucleotides (8-16 amino acids), wherein the percentage of sequence identity is calculated by comparing the reference sequence to the sequence having possible deletions or additions, which can be up to 20% of the reference sequence, and which are not included in the comparison window.

[0019] In the present application, the term "vagina" generally refers to the vaginal region or section or surrounding areas, including the labia, vulva, cervix, uterus, fallopian tubes, ovaries, urethra, bladder, anus, and rectum, including the mucosal tissue thereof. The term "reproductive tract" includes the vagina.

[0020] In the present application, the terms "disease" or "disorder" are used interchangeably and generally refer to any change in the state of the body or of some of its organs, impeding or disturbing the fulfillment of their functions, and / or causing symptoms such as discomfort, malfunction, suffering, or even death in a person suffering therefrom or in contact therewith.

[0021] In the present application, the term "pathogenic" (e.g., "pathogenic bacteria") generally refers to an agent, microorganism, or condition that is capable of causing a disease. In certain contexts, a pathogen also includes a microorganism (e.g., a bacterium) that is associated with a disease or disorder, but for which a causal relationship (e.g., a direct causal relationship) has not been established or is pending. In some embodiments, a microorganism that is not a pathogen and can be a commensal can cause or be associated with a disease or a dysbiosis, depending on various factors (e.g., the immune status of the site, the abundance of the microorganial taxon, etc.). Such a microorganism is referred to as a "pathobiont".

[0022] In the present application, the terms "pathogenic bacteria" or "pathogenic germs" or "pathogen" are used interchangeably and generally refer to microorganisms (e.g. bacteria) that can be deposited in the urogenital tract and in particular to Gardnerella vaginalis, Prevotella, Neisseria gonorrhoeae Neisseria gonorrhoeae ), mycoplasmas, Mobiluncus, Candida albicans and Candida glabrata Candida glabrata , and to pathogens that can cause intestinal infections such as, for example, Enterobacteriaceae, including Escherichia coli Escherichia coli , Salmonella, Staphylococcus, Clostridium difficile Clostridium difficile and Shigella.

[0023] In the present application, the terms "vaginal flora" or "vaginal microbiota" are used interchangeably and generally refer to microorganisms that colonize the vagina.

[0024] In the present application, the terms "vaginal flora disorder" or "vaginal dysbiosis" generally refer to a disorder of the normal dynamics of the vaginal flora balance, such as a decrease in Gram-positive bacteria and / or an increase in Gram-negative bacteria compared to a healthy vaginal environment (e.g. a condition in which the vaginal mucosa is devoid of protective lactobacilli and is colonized by a large number of different non-lactobacilli, which can be symptomatic or asymptomatic). Specific disorders or dysbiosis include, but are not limited to, fungal (e.g. yeast infections), bacterial (e.g. bacterial vaginosis), viral or parasitic (e.g. trichomonas) infections. The infections include those characterized / diagnosed as vaginitis, vaginal candidiasis and vaginosis. Examples of some microorganisms that can trigger such infections include Candida, in particular Candida albicans Candida albicans ) and Candida tropicalis C. tropicalis ) and Candida glabrata C. glabrata , Gardnerella (vaginal), various mixed anaerobic bacteria and Peptostreptococcus bacteria. If left untreated, these vaginal infections can cause pathological secretion of the vagina, great discomfort for the female patient, and / or serious medical consequences.

[0025] In the present application, the term "inhibition" generally refers to the process of inhibiting or impeding the growth and reproduction of bacteria and their activity.

[0026] In the present application, the term "isolated" when applied to a nucleic acid or a protein generally means that the nucleic acid or protein is substantially free of other cellular components with which it is associated in nature. For example, it can be in a homogeneous state and can be in a dry solution or aqueous solution. Purity and homogeneity are typically determined using analytical chemistry techniques such as polyacrylamide gel electrophoresis or high performance liquid chromatography.

[0027] The term "isolated" when applied to a bacterium generally refers to a bacterium that has been (1) separated from at least some of the components with which it is associated in nature (whether in the environment or in an experimental setting), and / or (2) produced, manufactured, purified, and / or manufactured by the hand of man, for example using artificial culture conditions such as, but not limited to, culturing on plates and / or in fermentors. Isolated bacteria include those that are cultured, even if such cultures are not single cultures. Isolated bacteria can be separated from at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or more of the other components with which it was originally associated. In embodiments, isolated bacteria are greater than about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or greater than about 99% pure. In certain embodiments, the populations of bacteria provided herein include isolated bacteria. In certain embodiments, The compositions provided herein include isolated bacteria. In certain embodiments, the bacteria administered are isolated bacteria.

[0028] In the present application, the term "pharmaceutically acceptable carrier" generally refers to a material that aids in the administration of an active agent to a subject and is absorbed by the subject and can be included in the compositions of the present application without causing a significant adverse toxicological effect on the patient. Non-limiting examples of pharmaceutically acceptable carriers include water, NaCl, normal saline solutions, lactated Ringer's, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavorants, salt solutions (such as Ringer's solution), alcohols, oils, gelatins, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethyl cellulose, polyvinyl pyrrolidine, and dyes, etc. Such preparations can be sterilized, and those of skill in the art will recognize that other pharmaceutical carriers can be useful in the present application.

[0029] In the present application, the term "CFU (Colony-Forming Units)" generally refers to the total number of colonies of bacteria, fungi, yeast, etc. microorganisms in a product, often used as a calculation of the number of viable bacteria.

[0030] The term "CFU / dose" means the amount of bacteria present in the composition / food or dietary supplement / medicament provided to the subject per day or per administration. For example, in certain embodiments, L. gasseri is present in the food or dietary supplement in an amount of from 10 6 to 10 12 CFU / dose (e.g., 10 8 to 10 12 CFU / dose). In such embodiments, if L. gasseri is administered in a food product (e.g., in a solid beverage, yogurt), the food product (e.g., solid beverage, yogurt) provided to the subject per day or per administration can contain about 10 6 to 10 12 CFU of L. gasseri. Of course, alternatively, the amount of such bacteria can be divided into multiple administrations, so long as the total amount of L. gasseri received by the subject in any particular time (e.g., 24 hour period) is from about 10 6 to about 10 12 CFU of bacteria, i.e., so long as L. gasseri is present in the food product or dietary supplement in an amount of from 10 6 to 10 12 CFU / dose (e.g., 10 8 to 10 12 CFU / dose).

[0031] In the present application, the term "treatment and / or prevention" includes not only treatment and / or prevention of a disease, but also generally includes preventing the onset of the disease, slowing or reversing the course of the disease, preventing or slowing the onset of one or more symptoms associated with the disease, reducing and / or alleviating one or more symptoms associated with the disease, reducing the severity and / or duration of the disease and / or any symptoms associated therewith, and / or preventing further increases in the severity of the disease and / or any symptoms associated therewith, preventing, reducing, or reversing any physiological damage caused by the disease and any pharmacological effects generally associated with treating the patient that are beneficial. The compositions of the present application form viable therapeutic agents without requiring complete cure or eradication of any symptoms or manifestations of the disease. As is recognized in the relevant art, a drug used as a therapeutic agent can reduce the severity of a given disease state, but need not eliminate every manifestation of the disease to be considered a useful therapeutic agent. Similarly, a therapeutic agent prophylactically administered forms a viable prophylactic agent without being completely effective in preventing the onset of the disorder. It is sufficient simply to reduce the impact of the disease in the subject (e.g., by reducing the number or severity of its symptoms, or by increasing the effectiveness of another therapy, or by producing another beneficial effect), or to reduce the likelihood of the disease occurring or worsening.

[0032] In the present application, the term "about" generally means a variation within a range of about 0.5-10% above or below the specified value, for example, a variation within a range of about 0.5%, about 1%, about 1.5%, about 2%, about 2.5%, about 3%, about 3.5%, about 4%, about 4.5%, about 5%, about 5.5%, about 6%, about 6.5%, about 7%, about 7.5%, about 8%, about 8.5%, about 9%, about 9.5%, or about 10% above or below the specified value.

[0033] Bacterial strain The Lactobacillus gasseri WT002 provided in the present application is derived from the genital tract secretion of a healthy married woman, and is identified as Lactobacillus gasseri by 16S rDNA Lactobacillus gasseri The strain is gram-positive, and the colony under microscope is white and opaque, smooth, with a white protrusion in the center, round, and with irregular edges, and the colony diameter is about 0.4-1.4 mm.

[0034] Lactobacillus gasseri Lactobacillus gasseri WT002, the preservation unit: China General Microbiological Culture Collection Center (CGMCC for short), address: No. 3, Beizhanxi Road, Chaoyang District, Beijing, preservation date: February 18, 2025, preservation number: CGMCC No. 33552.

[0035] In the present application, "Lactobacillus gasseri Lactobacillus gasseri WT002", "Lactobacillus gasseri WT002", "WT002", and "strain WT002" are synonymous.

[0036] The Lactobacillus gasseri of the present application has the advantages of strong growth vigor, low pH tolerance, high lactic acid production, antibacterial activity, and strong vaginal cell adhesion capacity.

[0037] Fermentation supernatant The present application provides a fermentation supernatant. The fermentation supernatant contains the aforementioned Lactobacillus gasseri Lactobacillus gasseri WT002 and / or the aforementioned metabolic product of Lactobacillus gasseri Lactobacillus gasseri WT002. It should be noted that "fermentation supernatant" refers to the supernatant after centrifugation of the fermentation broth, which may contain Lactobacillus gasseri WT002, or metabolic products of Lactobacillus gasseri WT002, or both Lactobacillus gasseri WT002 and its metabolic products. The fermentation supernatant of Lactobacillus gasseri WT002 of the present application has the advantage of promoting the proliferation of vaginal beneficial bacteria.

[0038] Composition The present application provides a composition containing the aforementioned Lactobacillus gasseri Lactobacillus gasseri) WT002 and / or the fermentation supernatant of the foregoing, and optionally one or more pharmaceutically acceptable carriers. The compositions of the present application are used for the prevention and / or treatment of diseases associated with vaginal dysbiosis.

[0039] In the present application, "pharmaceutically acceptable" means that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients of the formulation and / or the mammal being treated with it. Preferably, "pharmaceutically acceptable" as used herein means approved or approvable by a regulatory agency of the Federal or state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

[0040] In the present application, the term "pharmaceutically acceptable carrier" generally refers to a substance that aids in the administration of an active agent to a subject and is absorbed by the subject and can be included in the compositions of the present application without

[0041] In the present application, the term "pharmaceutically acceptable excipient" can include any solvent, suitable for the particular dosage form of interest. Except to the extent that any conventional excipient is incompatible with Lactobacillus gasseri WT002 as disclosed herein, for example, produces any adverse biological effect or interacts in a deleterious manner with any other component(s) of the pharmaceutically acceptable composition, their use is contemplated to be within the scope of this disclosure.

[0042] It is to be understood that the compositions of the present application need not achieve a complete cure or eradication of any symptom or manifestation of a disease to form a useful therapeutic agent. As is recognized in the relevant art, a drug used as a therapeutic agent can reduce the severity of a given disease state, but need not eliminate every manifestation of the disease to be considered a useful therapeutic agent. Similarly, a therapeutic agent administered prophylactically need not be completely effective in preventing the onset of a disorder to form a useful prophylactic agent. It is sufficient to simply reduce the impact of the disease in a subject (e.g., by reducing the number or severity of its symptoms, or by increasing the effectiveness of another therapy, or by producing another beneficial effect), or to reduce the likelihood of the disease occurring or worsening.

[0043] Uses The present application provides a Lactobacillus gasseri as previously described Lactobacillus gasseri) WT002, the aforementioned fermentation supernatant or the aforementioned composition for use in the preparation of a product for preventing and / or treating diseases associated with vaginal flora disorders caused by the proliferation of pathogenic bacteria, or for the adhesion and colonization of vaginal epithelial cells and human cervical cancer cells.

[0044] As a specific embodiment, the pathogenic bacteria include, but are not limited to, Escherichia coli, Gardnerella, Candida albicans and Prevotella.

[0045] As a specific embodiment, the diseases associated with vaginal flora disorders are reproductive tract infections. As a specific embodiment, the reproductive tract infections include, but are not limited to, bacterial vaginitis, fungal vaginitis, trichomonas vaginitis, aerobic vaginitis, senile vaginitis and vaginitis caused by viral infections. As a specific embodiment, the product is a food, a drug, a health product or a sanitary product.

[0046] As a specific embodiment, the food includes, but is not limited to, probiotic yogurt, probiotic tablets and probiotic solid beverages.

[0047] As a specific embodiment, the sanitary product can be a medical device, such as a colposcope, a gynecological self-checking mirror, a guide dilator, a speculum, a gynecological flusher or an antibacterial device for external use by women. In some embodiments, the sanitary product can also include a lotion, a panty liner, a paper towel, a tampon, a gel, an ointment, a cream, a spray or a shower gel.

[0048] The Lactobacillus gasseri WT002 and its fermentation supernatant disclosed in the present application can efficiently inhibit vaginal pathogenic bacteria, repair the microecological balance, and prevent the recurrence of reproductive tract infections by adhering to and colonizing vaginal epithelial cells and human cervical cancer cells, thus having significant advantages.

[0049] The solutions of the present application will be explained below in conjunction with examples. Those skilled in the art will understand that the following examples are only used to illustrate the present application and should not be regarded as limiting the scope of the present application. If the specific techniques or conditions are not specified in the examples, the techniques or conditions described in the literature in the art or according to the product instructions are used. If the manufacturers of the reagents, probiotics, pathogenic bacteria or instruments are not specified, they are all conventional products that can be obtained from the market.

[0050] Example 1: Isolation and identification of Lactobacillus gasseri WT002 Isolation of Lactobacillus gasseri WT002 The collected vaginal secretion samples of healthy married women were soaked in sterile normal saline, then 0.2 g of the sample was taken in 1 mL of sterile PBS (PBS stands for Phosphate-Buffered Saline) in an anaerobic operation box, and after being fully shaken and mixed, gradient dilution was performed and coated on PYG medium (PYG medium stands for Peptone-Yeast Extract-Glucose Medium), and the coated PYG medium plate was placed in a 37°C anaerobic condition (the gas composition of anaerobic is N2:CO2:H2=90:5:5) for 3 days, then single colonies were picked and streaked for isolation, and pure culture strains of each single strain were obtained, numbered as WT002.

[0051] Identification of Lactobacillus gasseri WT002 1) Morphological observation After the above-mentioned isolated strain WT002 was inoculated on MRS solid medium (MRS stands for de Man, Rogosa and Sharpe Agar) and cultured at 37°C under anaerobic conditions (the gas composition of anaerobic is N2:CO2:H2=90:5:5) for 48 hours, the colony morphology was observed by naked eye, and the colony of Lactobacillus gasseri WT002 was white and opaque, smooth, with a white protrusion in the center, round, and the edge was irregular, with a diameter of about 0.4-1.4 mm.

[0052] 2) Gram staining microscopy The above-mentioned strain WT002 single colony was picked for Gram staining, the slide was placed under the optical microscope, and after observation with a low-power lens, a drop of turpentine (or oil) was dropped on the slide, and the morphology and color of the bacteria were observed with a 1000-power oil lens. According to the experimental results, the WT002 bacterial body was slender rod-shaped, and the Gram staining was positive, without spores and flagella.

[0053] 3) Molecular biology identification 16S rDNA gene sequence analysis: The strain WT002 obtained by the above separation was cultured in liquid PYG medium at 37°C under anaerobic conditions for one day, 1 mL of bacterial liquid was centrifuged at 10000 r / min for 5 min, the bacterial body was collected, the genomic DNA was extracted, and the extracted genomic DNA was used as a template to perform PCR amplification using 16S rDNA universal primers 27F and 1492R. Primer 27F (sequence: 5'-AGAGTTTGATCMTGGCTCAG-3') is recorded as SEQ ID NO: 2, and amplification starts from the front end (5' end) of the 16S gene. Primer 1492R (sequence: 5'-GGTTACCTTGTTACGACTT-3') is recorded as SEQ ID NO: 3, and amplification starts from the end (3' end) of the 16S gene. The two primers are combined to amplify, and the conserved region of the 16S rDNA gene is amplified and sequenced. The obtained 16S rDNA amplification product is subjected to electrophoresis detection, purification, and 3730 sequencing, so as to obtain a 16S rDNA sequence with a length of about 1546 bp.

[0054] The 16S rDNA sequence obtained after sequencing the above PCR product is subjected to blast comparison analysis in GenBank. After homology comparison (BLASTN) with the standard sequences published in the GenBank database, it is found that the strain of WT002 has the highest similarity with Lactobacillus gasseri in the GenBank database, and the similarity is 100%. It is preliminarily confirmed that WT002 belongs to Lactobacillus gasseri.

[0055] According to the results of morphology, gram staining and molecular biology identification, the isolated WT002 strain is Lactobacillus gasseri ( Lactobacillus gasseri ), which was preserved in the China General Microbiological Culture Collection Center on February 18, 2025, at No. 1, Beichen West Road, Chaoyang District, Beijing, with a preservation number of CGMCC NO. 33552.

[0056] The 16S rDNA sequencing result of Lactobacillus gasseri WT002 is shown in SEQ ID NO: 1.

[0057]

[0058] Example 2: Physiological and biochemical characteristics of Lactobacillus gasseri WT002 1) Growth curve of Lactobacillus gasseri WT002 The isolated strain WT002 was inoculated into MRS medium and cultured at 37°C under anaerobic conditions for 24 hours to an OD600 value of about 0.8-1, and 1% of Lactobacillus gasseri WT002 was inoculated into new MRS medium at a volume ratio, and continued to be cultured in a constant temperature shaker at 37°C. Every 2 hours, sample and measure OD600 value. The growth curve was drawn by the measured data, and the results are shown in Figure 1 As shown in the figure, the WT002 strain showed exponential growth phase within 0 to 12 hours, and tended to plateau after 12 hours, indicating that the growth of the strain WT002 in MRS medium had good growth ability.

[0059] 2) Catalase reaction detection The isolated strain WT002 was inoculated into MRS medium and cultured at 37°C under anaerobic conditions for 48 hours, and single colonies were picked for catalase reaction detection. According to the experimental results, the catalase reaction of Lactobacillus gasseri WT002 was positive.

[0060] It can be understood that catalase can neutralize the bactericidal effect of hydrogen peroxide (H2O2) by decomposing hydrogen peroxide (H2O2) in vivo. In other words, the Lactobacillus gasseri WT002 provided by the present application has the effect of regulating the amount of hydrogen peroxide produced.

[0061] 3) Sugar alcohol fermentation detection The isolated strain WT002 was inoculated into MRS medium and cultured at 37°C under anaerobic conditions for 48 hours, and single colonies were picked for sugar alcohol fermentation detection. Different types of sugar alcohols (such as sorbitol, mannitol, etc.) were used as fermentation substrates in the experiment. After the culture was completed, the pH change in the culture medium was monitored, and the presence or absence of gas production was detected by the Durham tube to determine whether the strain could effectively ferment sugar alcohol. The fermentation capacity and end product were further analyzed by measuring the OD value and acid production. According to the experimental results, the WT002 sugar alcohol fermentation detection was positive, and the strain was facultative anaerobic, which could effectively metabolize sugar alcohol under anoxic environment.

[0062] 4) Carbon source utilization detection The isolated strain WT002 was inoculated into MRS medium and cultured at 37°C under anaerobic conditions for 48 hours, and single colonies were picked for carbon source utilization detection, and the results are shown in Table 1: Table 1 Carbon source utilization of Lactobacillus gasseri WT002

[0063] Note: + is positive, - is negative, W is weakly positive.

[0064] As can be seen from Table 1, the Lactobacillus gasseri WT002 of the present application can efficiently utilize monosaccharides (such as D-glucose, D-galactose, D-fructose, D-mannose, D-tagatose) and disaccharides (such as D-maltose, D-sucrose, D-lactose, D-cellulobiose, D-trehalose, D-melibiose, D-gentiobiose), indicating that the strain can quickly utilize common monosaccharides and dietary disaccharides. Since monosaccharides are one of the main carbon sources in the intestinal tract, Lactobacillus gasseri WT002 can adapt to the intestinal environment and help in scenarios such as lactose intolerance. Moreover, Lactobacillus gasseri WT002 can also utilize trisaccharides (such as D-melibiulose, D-raffinose) and fructans (such as inulin) Further, Lactobacillus gasseri WT002 can also decompose starch, N-acetylglucosamine (mucin component), aescin, salicin, etc., indicating that Lactobacillus gasseri WT002 can degrade complex carbohydrates, and its ability to utilize N-acetylglucosamine (mucin component) can help enhance intestinal adhesion or colonization ability.

[0065] In addition, Lactobacillus gasseri WT002 has low utilization of sugar alcohol carbon sources (such as L-sorbitol), which can reduce the negative impact of sugar alcohol intolerance.

[0066] It can be understood that the WT002 strain can utilize various forms of carbon sources, has a wide carbon source metabolic capacity, can compete for carbon sources in complex environments, and has a survival advantage. This metabolic characteristic makes it a potential multifunctional intestinal probiotic, which can be applied in the food industry, especially in the development of probiotic products, and has certain market application potential.

[0067] 5) Lactic acid production ability detection Inoculate the strain WT002 into MRS medium, and cultivate at 37°C for 24 h under anaerobic and aerobic conditions, respectively. Centrifuge 1 mL of bacterial liquid at 8000 r / min for 5 min, and take the supernatant as the mother liquor of the test sample.

[0068] Use commercially available L-Lactic Acid (L-Lactate) Assay Kit and D-Lactic Acid (D-Lactate) Assay Kit (manufacturer: Sigma-Aldrich) respectively and follow the instructions to process the samples as shown in Table 2, as the test sample solution.

[0069] (1) The detection of L-lactic acid production is shown in Table 3.

[0070] After adding 0.02 mL of suspension 4 (D-LDH) to the blank control group and the experimental group and mixing thoroughly, the absorbance A1 of the blank control group and the absorbance A2 of the experimental group were measured at 340 nm using an ultraviolet spectrophotometer. The absorbance was measured every 1 minute for 5 minutes, until the absorbance remained unchanged, and the L-lactic acid concentration was calculated according to formula (1): C (g / L) = 0.3204 x ΔA (L-lactic acid), ΔA = (A2-A1).

[0071] (2) Detection of D-lactic acid production, and the results are shown in Table 3.

[0072] After adding 0.02 mL of suspension 4 (D-LDH) to the blank control group and the experimental group and mixing thoroughly, the absorbance A1 of the blank control group and the absorbance A2 of the experimental group were measured at 340 nm using an ultraviolet spectrophotometer. The absorbance was measured every 1 minute for 5 minutes, until the absorbance remained unchanged, and the L-lactic acid concentration was calculated according to formula (1): C (g / L) = 0.3204 x ΔA (L-lactic acid), ΔA = (A2-A1).

[0073] Table 2

[0074] Table 3

[0075] It should be understood that the lactic acid produced by the lactobacillus in the reproductive tract can inhibit the growth of pathogenic bacteria. As can be seen from Table 3, the L-lactic acid and D-lactic acid produced by the Lactobacillus gasseri WT002 obtained in the application under anaerobic and aerobic conditions is high, indicating that the Lactobacillus gasseri WT002 provided in the application can produce lactic acid under both environments, i.e., has strong bacteriostatic potential, and can be used as a vaginal microecological regulator to maintain human vaginal health.

[0076] 6) Detection of hydrogen peroxide production Strain WT002 was inoculated into MRS medium and cultured at 37°C for 24 h under aerobic and anaerobic conditions, respectively. 1 mL of bacterial solution was added with an appropriate amount of lysozyme (lysozyme final concentration was 1 mg / mL) to lyse the cell wall of WT002 strain, and release hydrogen peroxide in the solution. After standing at 37°C for 15 min, centrifugation was performed at 8000 r / min for 5 min, and the supernatant was used as the test solution. The content of hydrogen peroxide in the test solution was detected using a commercially available hydrogen peroxide detection kit (manufacturer: Sigma-Aldrich), and the results are shown in Table 4.

[0077] Table 4

[0078] It should be understood that hydrogen peroxide (i.e., H2O2) can inhibit the growth of pathogenic bacteria, and the Lactobacillus gasseri WT002 provided in the present application can produce hydrogen peroxide under anaerobic and aerobic conditions, and play a probiotic role. And because the catalase reaction of Lactobacillus gasseri WT002 is positive, it not only has excellent hydrogen peroxide production ability, but also can participate in the regulation of H2O2 level in the organism, thereby enhancing its own colonization ability, inhibiting pathogenic bacteria, and maintaining the balance of microecology.

[0079] 7) Test of tolerance to acid and bile salts (1) Acid tolerance test Prepare MRS culture medium with pH=2, pH=3, pH=4, pH=4.5, and pH=7, respectively, and inoculate 100 μL of overnight culture of WT002 bacterial solution (concentration of 2.0E+08 CFU / mL) into the MRS culture medium with different pH values, and then incubate at 37°C under anaerobic conditions for 24 h. Plate the bacterial solution and count the colonies, and the results are shown in Table 5.

[0080] Table 5: Acid tolerance of WT002

[0081] As can be seen from Table 5, Lactobacillus gasseri WT002 can survive and grow under the conditions of pH=2, pH=3, pH=4, pH=4.5, and pH=7.

[0082] It can be understood that the pH value in the normal female vaginal environment is between 3.8-4.4, and the Lactobacillus gasseri WT002 provided in the present application has low-pH growth activity, and has excellent growth activity in an acidic environment with pH=4.5. Further, when the vaginal flora is disordered and other pathogenic bacteria dominated by aerobic bacteria proliferate excessively, resulting in an increase in the pH of the vagina, the Lactobacillus gasseri WT002 still has excellent growth activity, which indicates that the Lactobacillus gasseri WT002 can adapt to the complex reproductive tract environment, ensure long-term colonization in the reproductive tract to provide continuous protection, form niche occupation, and quickly restore the acidic environment through its metabolic products to inhibit the growth of pathogenic bacteria and reduce the recurrence rate of vaginal infections.

[0083] (2) Choline tolerance test Prepare MRS culture medium containing 0.05%, 0.10%, 0.20%, and 0.30% bile salts, respectively, and inoculate 100 μL of overnight culture of WT002 bacterial solution (concentration of 2.5E+08 CFU / mL) into the MRS culture medium with different bile salt contents, and then incubate at 37°C under anaerobic conditions for 24 h. Plate the bacterial solution and count the colonies, and the results are shown in Table 6.

[0084] Table 6 Cholate salt tolerance of WT002

[0085] It should be understood that cholate salt is the main component of bile, which can destroy the microbial cell membrane and thus kill intestinal pathogens. The normal healthy human small intestine cholate salt concentration is usually 0.03%-0.30%. As can be seen from Table 6, Lactobacillus gasseri WT002 has cholate salt tolerance, which can survive and grow under the conditions of 0.05%, 0.10%, 0.20%, and 0.30% cholate salt, which means that Lactobacillus gasseri WT002 can pass through the gastric acid and duodenal cholate salt environment, survive to the intestinal tract and colonize, that is, it can be applied as an oral probiotic to compete with pathogenic bacteria in the intestinal tract.

[0086] 8) Antibiotic sensitivity test Strain WT002 was inoculated into MRS liquid medium, cultured under anaerobic conditions at 37°C for 24 h, 0.1 ml of bacterial liquid was spread on MRS solid medium, and antibiotic drug sensitive sheets to be detected (ampicillin, bacitracin, penicillin, kanamycin, tetracycline, amoxicillin, erythromycin, chloramphenicol, ceftriaxone, vancomycin, oxacillin, amoxicillin, azithromycin, clindamycin and gentamicin) were pasted on the surface of the plate, and the plate was cultured at 37°C for 24 h. The diameter of the inhibition zone was measured, and the test results are shown in Table 7.

[0087] Table 7 Antibiotic resistance of Lactobacillus gasseri WT002

[0088] As can be seen from the results of Table 7, the Lactobacillus gasseri WT002 provided by the present application is resistant to bacitracin, clindamycin and gentamicin, and is sensitive to ampicillin, penicillin, kanamycin, tetracycline, amoxicillin, erythromycin, chloramphenicol, ceftriaxone, vancomycin, oxacillin, amoxicillin and azithromycin, and has no drug resistance, and can be safely used. The drug resistance spectrum shows that Lactobacillus gasseri WT002 is suitable for use as a probiotic.

[0089] Example 3: Inhibition ability test of Lactobacillus gasseri WT002 on four kinds of vaginal pathogenic bacteria 1) Inhibition of Escherichia coli E. coli ATCC35218 test 2.5 μL of overnight culture of Lactobacillus gasseri WT002 was spotted on MRS solid medium, and the plate was cultured under anaerobic conditions at 37°C for 24 h to obtain a bacterial plaque of Lactobacillus gasseri WT002.

[0090] 500 μL of 10 7 CFU / mL of Escherichia coli E. coliATCC35218 was mixed with 10 ml of LB semi-solid medium (LB stands for Luria-Bertani Medium) containing 0.5% agar. The mixture was then poured onto a plate inoculated with WT002 plaques. After the medium solidified, it was incubated at 37°C for 24 h under anaerobic conditions. This was the test group. The diameter of the inhibition zone was observed and recorded. The results are shown in Table 8.

[0091] Lactobacillus gasseri GDMCC 60092 was used as the control group. The culture method of the control group was the same as that of the test group, and the size of the inhibition zone diameter was observed and recorded. The results are shown in Table 8.

[0092] 2) Inhibit Gardnerella vaginalis Gardnerella vaginalis BNCC337545 test 2.5 μL of overnight cultured Lactobacillus gasseri WT002 was inoculated onto MRS solid medium and cultured under anaerobic conditions at 37°C for 24 h to obtain Lactobacillus gasseri WT002 plaques.

[0093] Take 500 μL 10 7 Gardnerella vaginalis CFU / mL Gardnerella vaginalis BNCC337545) was mixed with 10 mL of Gardnerella Columbia blood agar semi-solid medium containing 0.5% agar. The mixture was then poured onto a plate inoculated with WT002 plaques. After the medium solidified, it was incubated at 37°C for 24 h under anaerobic conditions. This was the test group. The diameter of the inhibition zone was observed and recorded. The results are shown in Table 8.

[0094] Lactobacillus gasseri GDMCC 60092 was used as the control group. The culture method of the control group was the same as that of the test group, and the size of the inhibition zone diameter was observed and recorded. The results are shown in Table 8.

[0095] Table 8. Inhibitory effect of Lactobacillus gasseri WT002 on Escherichia coli and Gardnerella vaginalis.

[0096] As shown in Table 8, in the Escherichia coli inhibition test, the inhibition zone diameter of the control group Lactobacillus gasseri GDMCC 60092 was 0.91±0.3 cm, which was smaller than the inhibition zone diameter of Lactobacillus gasseri WT002 (1.48±0.15 cm) provided in this application. This means that the Lactobacillus gasseri WT002 provided in this application has a lower inhibition zone diameter than Escherichia coli. E. coli ATCC35218 showed higher inhibitory activity than Lactobacillus gasseri GDMCC 60092.

[0097] Furthermore, in the Gardnerella vaginalis inhibition test, the inhibition zone diameter of the control group Lactobacillus gasseri GDMCC 60092 was 0.61±0.18 cm, which was smaller than the inhibition zone diameter of Lactobacillus gasseri WT002 of this application (0.83±0.16 cm). This indicates that the inhibitory ability of Lactobacillus gasseri WT002 provided in this application against Gardnerella vaginalis is higher than that of Lactobacillus gasseri GDMCC 60092.

[0098] 3) Inhibits Candida albicans Candida albicans SC5314 test Overnight activated Lactobacillus gasseri WT002 and Candida albicans ( Candida albicans SC5314 was co-inoculated into a modified PYG medium (prepared from 5 g tryptone, 3 g yeast extract, 10 g glucose, 0.5 g tyrosine, 2 g KH2PO4, 1 g K2HPO4, 0.2 g MgSO4·7H2O, 0.1 g CaCl2, and 15 g agar) and cultured under anaerobic conditions at 37°C for 24 h to obtain a mixed bacterial culture.

[0099] The above mixed bacterial suspension was diluted and spread onto PDA fungal culture plates (PAD stands for Potato Dextrose Agar Plate). After incubation at 37°C for 24 hours under aerobic conditions, this was used as the test group. Correspondingly, negative and positive control groups were set up based on the conditions of the test group. The difference between the negative and positive control groups and the test group was that the negative control group was inoculated only with *Lactobacillus gasseri* WT002, and the positive control group was inoculated only with *Candida albicans*. Candida albicans (SC5314) Colony counts were performed on the test group, negative control group, and positive control group. The inhibition rate of the test group was calculated according to the formula: inhibition rate (%) = (colony count in positive control group - colony count in test group) / colony count in positive control group × 100%. The results are shown in Table 9.

[0100] 4) Inhibits Prevotella Prevotella bivia ATCC 29563 test Overnight activated Lactobacillus gasseri WT002 and Prevotella ( Prevotella bivia ATCC 29563) was co-inoculated into modified PYG medium and cultured under anaerobic conditions at 37°C for 24 hours to obtain a mixed bacterial culture.

[0101] The above mixed bacterial solution was diluted and spread onto PDA fungal culture plates. After incubation at 37°C for 24 hours under aerobic conditions, it served as the test group. A negative control group and a positive control group were set up accordingly. The negative control group and the positive control group differed from the test group in that the negative control group was inoculated only with *Lactobacillus gasseri* WT002, and the positive control group was inoculated only with *Prevotella*. Prevotella bivia According to ATCC 29563, colony counts were performed on the test group, negative control group, and positive control group, and the inhibition rate (%) was calculated using the formula: Inhibition rate (%) = (Number of colonies in positive control group - Number of colonies in test group) / Number of colonies in positive control group × 100%. The inhibition rate of the test group was calculated, and the results are shown in Table 9.

[0102] Table 9. Inhibitory effect of Lactobacillus gasseri WT002 on Candida albicans and Prevotella.

[0103] As shown in Table 9, the Lactobacillus gasseri WT002 provided in this application has a significant inhibitory effect on the growth of Candida albicans and Prevotella. According to the experimental results, the inhibition rate of Lactobacillus gasseri WT002 against Candida albicans is 99.39%, and the inhibition rate against Prevotella is as high as 99.68%.

[0104] Example 4: Adhesion assay of human cervical cancer cells HeLa and human vaginal epithelial cells VK2E6 / E7 Cell processing: Cultured human cervical cancer cells (HeLa) and human vaginal epithelial cells (VK2E6 / E7, purchased from Beijing Beina Chuanglian Biotechnology Research Institute) were digested separately and diluted with 1640 complete culture medium (GIBCO, purchased from Wokaway Beijing Biotechnology Co., Ltd.) without antibiotics to a cell concentration of approximately 2 × 10⁻⁶ cells / year. 5 After counting cells / mL (using a hemocytometer), take 1 mL of cell solution into a cell culture dish (12-well or 6-well plate) and incubate at 37°C in a 5% CO2-95% air incubator until complete differentiation. After the cells grow into a dense monolayer, wash the cells twice with sterile PBS buffer.

[0105] (1) HeLa test for adhesion of human cervical cancer cells Add 1 mL of 1640 culture medium and 1 mL of Lactobacillus gasseri WT002 bacterial suspension (obtained by anaerobic culture at 37°C for 24 h) to each well of the above-mentioned cell culture dish containing HeLa cells, and incubate overnight in an air incubator, adjusting the temperature to 10. 8 The bacterial suspension was gently shaken and then incubated in an incubator at 37°C and 5% CO2. Each bacterial sample was replicated in three wells.

[0106] After 90 min of incubation, the cell culture dish was removed, the bacterial suspension was discarded, and the cells were washed 5 times with sterilized PBS buffer to remove the unadhered lactobacilli. Anhydrous methanol was added to the cell culture dish for fixation for 20 min. The cell glass slide fixed with anhydrous methanol was removed and subjected to Gram staining as the experimental group. The Lactobacillus gasseri GDMCC 60092 was used as the control group. The control group was treated in the same manner as the experimental group, except that the Lactobacillus gasseri WT002 bacterial solution was replaced with the Lactobacillus gasseri GDMCC 60092. The cell glass slides of the experimental group and the control group were dried and observed under a microscope for counting. The number of bacteria adhered to 100 cells in 30 random fields was calculated, and the results are shown in Table 10.

[0107] (2) VK2E6 / E7 human vaginal epithelial cell adhesion test In the cell culture dish containing the VK2E6 / E7 human vaginal epithelial cells, 1 mL of 1640 culture solution and 1 mL of the Lactobacillus gasseri WT002 bacterial solution (obtained by culturing under anaerobic conditions at 37°C for 24 h) were added. The air incubator was used for overnight culture, and the number of bacteria was adjusted to 10 8 After gentle shaking, incubation was continued at 37°C in a 5% CO2 incubator, and three wells were repeated for each strain.

[0108] After 90 min of incubation, the cell culture dish was removed, the bacterial suspension was discarded, and the cells were washed 5 times with sterilized PBS buffer to remove the unadhered lactobacilli. Anhydrous methanol was added to the cell culture dish for fixation for 20 min. The cell glass slide fixed with anhydrous methanol was removed and subjected to Gram staining as the experimental group. The Lactobacillus gasseri GDMCC 60092 was used as the control group. The control group was treated in the same manner as the experimental group, except that the Lactobacillus gasseri WT002 bacterial solution was replaced with the Lactobacillus gasseri GDMCC 60092. The cell glass slides of the experimental group and the control group were dried and observed under a microscope for counting. The number of bacteria adhered to 100 cells in 30 random fields was calculated, and the results are shown in Table 10.

[0109] Table 10

[0110] As shown in Table 10, each Hela cell can adhere to 22.35 ± 1.79 Lactobacillus gasseri GDMCC 60092, and each Hela cell can adhere to 55.13 ± 2.75 Lactobacillus gasseri WT002. That is, the adhesion ability of the Lactobacillus gasseri WT002 to the human cervical cancer cell Hela is higher than that of the control group Lactobacillus gasseri GDMCC 60092.

[0111] And, each human vaginal epithelial cell VK2E6 / E7 adhered an average of 41.03 ± 3.66 Lactobacillus gasseri GDMCC60092, while each human vaginal epithelial cell VK2E6 / E7 adhered an average of 126.17 ± 5.94 Lactobacillus gasseri WT002, in other words, the adhesion ability of Lactobacillus gasseri WT002 to human vaginal epithelial cells VK2E6 / E7 was higher than that of the control group Lactobacillus gasseri GDMCC60092.

[0112] It can be understood that the stronger the adhesion ability of Lactobacillus gasseri WT002 to human cervical cancer cells Hela and human vaginal epithelial cells VK2E6 / E7, the stronger the colonization ability of the strain, and the easier it is to survive in the reproductive tract environment for reproduction, thereby playing a role in inhibiting pathogenic bacteria, protecting the vaginal mucosa, and restoring the vaginal flora, achieving the therapeutic or prophylactic effect of treating or preventing vaginal infection symptoms.

[0113] Example 5: Test of Lactobacillus gasseri WT002 for promoting the proliferation of vaginal beneficial bacteria 1. Preparation of Lactobacillus gasseri WT002 fermentation product 2.5 μL of overnight culture of WT002 was anaerobically fermented in MRS liquid medium at 37°C for 24 h, centrifuged at 8000 x g for 10 min, and the supernatant was obtained. The supernatant was sterilized with a 0.22 μm filter membrane to obtain the WT002 fermentation product.

[0114] 2. Collection and culture of vaginal beneficial bacteria (1) Lactobacillus crispatus Lactobacillus crispatus ATCC 33820: Lactobacillus crispatus was picked Lactobacillus crispatus ATCC 33820 single colony was inoculated in MRS broth medium and cultured at 37°C under anaerobic conditions for 24 h, and the culture was diluted to about 5 x 10 5 CFU / mL.

[0115] (2) Lactobacillus iners Lactobacillus iners ATCC BAA-3226: Lactobacillus iners was taken from the Lactobacillus iners Lactobacillus iners ATCC BAA-3226 cryovial Lactobacillus iners ATCC BAA-3226, and inoculated in MRS broth medium with 0.05% L-cysteine, and cultured at 37°C under anaerobic conditions for 48 h, and the culture was adjusted to about 5 x 10 5 CFU / mL.

[0116] (3) Lactobacillus jensenii Lactobacillus jensenii ATCC 25258: Lactobacillus jensenii was picked Lactobacillus jenseniiSingle colonies of ATCC 25258 were inoculated into MRS broth medium and incubated statically at 37°C under anaerobic conditions for 48 h to ensure that the strain reached a suitable growth state.

[0117] 3. Test on the effect of WT002 fermentation products on the proliferation of beneficial vaginal bacteria (1) Promotes the growth of Lactobacillus curvature Lactobacillus crispatus ATCC 33820 proliferation assay In 96-well plates, 180 μL of fresh MRS liquid culture medium and 20 μL of WT002 fermentation product were added to each well of the experimental group, while an equal volume of sterile MRS was added to the control group. Both the experimental and control groups were inoculated with 20 μL of Lactobacillus flavus (…). Lactobacillus crispatus The experimental group (ATCC 33820) was cultured in an anaerobic incubator at 37℃ for 12 h. The OD600 was measured using a microplate reader. The OD600 gain of the experimental group and the control group was compared. The proliferation rate was calculated according to the formula. The experiment was repeated three times. The results are shown in Table 11.

[0118] Table 11. Fermentation products of WT002 promote the proliferation of Lactobacillus curvature.

[0119] The results showed that the fermentation products of strain WT002 had a significant promoting effect on the proliferation of Lactobacillus curvature, with a proliferation rate of 21.39% to 24.87%.

[0120] (2) Promotes the growth of inert lactobacilli Lactobacillus iners ATCC BAA-3226 Proliferation Assay 2.5 μL of WT002 fermentation product was inoculated at the center of an MRS plate and cultured anaerobically at 37°C for 24 h to form a WT002 metabolite diffusion zone. 100 μL of inert lactobacillus (…) was then added… Lactobacillus iners The ATCC BAA-3226 inoculum was mixed with 5 mL of 0.75% (w / v) MRS semi-solid medium (45–50℃), and then quickly poured onto and covered the surface of the plate where the WT002 metabolite diffusion zone had formed. After solidification, it was cultured under anaerobic conditions at 37℃ for 24 h. The inert Lactobacillus (…) was observed and measured. Lactobacillus iners The diameter of the proliferation zone formed in the ATCC BAA-3226 coating layer due to the WT002 fermentation product was used as the experimental group. The control group was treated the same as the experimental group, except that the control group was not inoculated with WT002 fermentation product. The two groups of inert Lactobacillus (ATCC BAA-3226) were compared. Lactobacillus iners The difference in the diameter of the proliferation zone of the ATCC BAA-3226 membrane layer was investigated, and the proliferation rate was calculated according to the formula. The experiment was repeated three times, and the results are shown in Table 12.

[0121] Table 12 WT002 fermentation product promotes the proliferation of Lactobacillus iners

[0122] The results show that the fermentation product of the WT002 strain has a significant promoting effect on the proliferation of Lactobacillus iners, and the proliferation promotion rate reaches 27.65% ~ 32.40%.

[0123] (3) Promoting the proliferation of Lactobacillus jensenii Lactobacillus jensenii ATCC 25258 proliferation test The experimental group took 4 mL of MRS liquid medium and mixed 1 mL of WT002 strain fermentation product into the culture tube, and the control group took the same volume of MRS liquid medium, the experimental group and the control group were inoculated with Lactobacillus jensenii at 1% inoculation amount, and cultured at 37°C under anaerobic conditions for 24 hours, the absorbance value at OD=600 nm was measured, and the results are shown in Table 13.

[0124] Table 13 WT002 fermentation product promotes the proliferation of Lactobacillus jensenii

[0125] The results show that the fermentation product of the WT002 strain has a significant promoting effect on the proliferation of Lactobacillus jensenii, and the proliferation promotion rate reaches 24.54% ~ 28.71%.

[0126] It should be understood that in the description of the present specification, the description of the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" and the like means that the specific features, structures, materials or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present application. In the present specification, the illustrative description of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any appropriate manner in any one or more embodiments or examples. In addition, different embodiments or examples described in the present specification and the features of different embodiments or examples can be combined and combined by those skilled in the art without contradiction.

[0127] The above-mentioned embodiment number of the present application is only for description, not representing the advantages and disadvantages of the embodiments. The above description is only a specific embodiment of the present application, but the protection scope of the present application is not limited thereto. Any person skilled in the art can easily think of various equivalent modifications or replacements within the technical scope disclosed by the present application, and these modifications or replacements should be included in the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

Claims

1. A type of Lactobacillus gasseri ( Lactobacillus gasseri WT002, characterized in that, The preservation number of the Lactobacillus gasseri is CGMCC No. 33552.

2. The use of Lactobacillus gasseri WT002 according to claim 1 in the preparation of a drug, characterized in that, The drug is used to prevent and / or treat reproductive tract infections.

3. The use of Lactobacillus gasseri WT002 according to claim 1 in the preparation of a drug, characterized in that, The drug is used to inhibit pathogenic bacteria.

4. The use according to claim 3, characterized in that, The pathogenic bacteria include at least one of Escherichia coli, Gardnerella vaginalis, Candida albicans, and Prevotella.

5. The use of Lactobacillus gasseri WT002 according to claim 1 in the preparation of a drug, characterized in that, The drug is used to prevent and / or treat reproductive tract flora imbalance.

6. The use of Lactobacillus gasseri WT002 according to claim 1 in the preparation of a drug, characterized in that, The drug has the function of vaginal epithelial cell adhesion.

7. The use of Lactobacillus gasseri WT002 according to claim 1 in the preparation of a drug, characterized in that, The drug has the function of adhering to human cervical cancer cells.

8. The use of Lactobacillus gasseri according to claim 1 in the preparation of a medicament, wherein the medicament is used in a product for improving reproductive tract flora, characterized in that, The improvement of reproductive tract flora includes promoting the proliferation of beneficial vaginal bacteria, which includes promoting the proliferation of Lactobacillus curvaturei (Lactobacillus curvaturei). Lactobacillus crispatus ) proliferation, promoting the growth of inert lactobacilli ( Lactobacillus iners ) proliferation and promotion of Lactobacillus janniae ( Lactobacillus jensenii )proliferation.

9. A composition, characterized in that, The composition comprises Lactobacillus gasseri WT002 as described in claim 1, and optionally one or more pharmaceutically acceptable carriers.

10. The use of Lactobacillus gasseri as described in claim 1 or the composition as described in claim 9 in the preparation of food, health products and hygiene products.

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