Witzia ciliaris and application thereof

Abstract

The invention relates to Witzia ciliaris and an application thereof. Specifically, the invention relates to Witzia ciliaris We06, and a composition, a culture, a food and a pharmaceutical composition containing the Witzia ciliaris We06. The invention also relates to application of the Witzia ciliaris We06 and a composition, a culture, a food and a pharmaceutical composition containing the Witzia ciliaris We06 in preparation of medicines.

Description

Technical Field

[0001] The present application relates to a coagulant Weizmannella and its application. Specifically, the present application relates to a coagulant Weizmannella We06 and a composition, culture, food and pharmaceutical composition comprising the same. The present application also relates to the use of the coagulant Weizmannella We06 and a composition, culture, food and pharmaceutical composition comprising the same in the preparation of medicines. Background Art

[0002] Lactic acid bacteria (LAB) are probiotics that can promote the decomposition of lipids, proteins and carbohydrates, improve the flavor of fermented products, and improve human health. They are widely used in markets such as functional foods and healthcare products. As of 2023, lactic acid bacteria include 25 genera, 589 species and 58 subspecies. They can use carbohydrates to produce organic acids and peptides to inhibit the growth of spoilage bacteria and pathogenic bacteria, and prevent the accumulation of heavy metals in different organs of the body. Currently, commonly used lactic acid bacteria include Lactobacillus casei, Lactococcus, and Leuconostoc, but traditional lactic acid bacteria are affected by their own biological characteristics, and there is a problem of decreased activity during the production, transportation and sales cycle of bacterial agents, resulting in reduced product effectiveness. Therefore, there is an urgent need to seek new alternative strains and improve the activity of bacterial agents.

[0003] Weizmannella coagulans is classified as a spore-forming lactic acid bacterium and is an important potential probiotic. Therefore, it is urgent to develop Weizmannella coagulans with good heat and acid resistance, which can maintain a relatively high survival rate after dairy products are sterilized or during the shelf life of dairy products and after consumption, so as to improve the efficacy of probiotic lactic acid bacteria in dairy products.

[0004] In addition, as a spore-forming lactic acid bacterium, the spore-forming characteristics of Weizmannella coagulans are also of great significance in improving the stability and safety of food and industrial products.

[0005] As a dormant form of bacteria, spores can resist the attack of high temperature, dryness, ultraviolet radiation and chemicals, allowing Weizmannella coagulans to survive in adverse environments and re-germinate and reproduce when conditions are suitable. This characteristic is particularly important in the food industry, as it can ensure that probiotic products survive high-temperature sterilization processes, thereby improving the stability and activity of the products. In addition, Weizmannella coagulans can be used to produce high-temperature resistant probiotic products, such as probiotic beverages and probiotic powders.

[0006] In agriculture, Weizmannella coagulans improves the disease resistance and growth ability of plants through its spore-forming ability, and is widely used as a biofertilizer and biopesticide. In industrial fermentation processes, its spore form can also be used to produce metabolites such as enzymes and antibiotics. Heat resistance not only makes Weizmannella coagulans suitable for food processing and industrial production processes with high temperature treatment, but also enables it to play an important role in environmental governance such as high-temperature composting and sludge treatment, promote the resource utilization of waste, and reduce the emission of harmful substances. By forming spores and possessing heat resistance, Weizmannella coagulans has shown broad application prospects and important value in many fields. Summary of the invention

[0007] After a large number of experiments, the inventors of the present application screened a strain of Weizmannella coagulans with significant probiotic potential from dairy products. Furthermore, the inventors of the present application confirmed through a large number of experiments that the Weizmannella coagulans has good tolerance to the acidic environment of gastric juice and bile salts, is heat-resistant, can adapt to the gastrointestinal environment, and has excellent spore production ability; and the Weizmannella coagulans is sensitive to antibiotics, does not have toxin genes, and has high food safety. Furthermore, the Weizmannella coagulans also has a certain inhibitory effect on pathogenic bacteria and fungi.

[0008] Therefore, in a first aspect, the present application provides a coagulant Weizmannella ( Weizmannia coagulans ), the coagulant Weizmannella is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number being CGMCC No. 28819.

[0009] In certain embodiments, the colonies of the Weizmannella coagulans are white, and the colony edges are not smooth and the surface is rough.

[0010] In certain embodiments, the Weizmannella coagulans is resistant to gastric acid and / or bile salts.

[0011] In certain embodiments, the Weizmannella coagulans is thermotolerant.

[0012] In certain embodiments, the Weizmannella coagulans has sporulation capability.

[0013] In certain embodiments, the Weizmannella coagulans is capable of inhibiting pathogenic bacteria and / or fungi.

[0014] In certain embodiments, the pathogenic bacteria is selected from Staphylococcus aureus ( Staphylococcus golden ), Escherichia coli ( Escherichia coli ), or any combination thereof.

[0015] In certain embodiments, the pathogenic fungus is Debaryomyces hansenii ( Debaryomyces hansenii).

[0016] In certain embodiments, the Weizmannella coagulans contains:

[0017] a) the nucleotide sequence shown in SEQ ID NO: 1; or,

[0018] b) a nucleotide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, at least 99.95%, at least 99.99% or 100% identical to the nucleotide sequence of SEQ ID NO: 1; or,

[0019] c) a complementary nucleotide sequence of a) or b).

[0020] In a second aspect, the present application provides a composition comprising the Weizmannella coagulans.

[0021] In certain embodiments, the Weizmannella coagulans may be used in combination with one or more other species of microorganisms that can have a beneficial effect on the health of the host to which it is administered. Thus, in certain embodiments, the composition further comprises additional probiotic bacteria and / or probiotic fungi (e.g., yeast), wherein the additional probiotic bacteria and / or probiotic fungi are edible.

[0022] As used herein, the term "probiotic bacteria / fungus" is defined as any non-pathogenic bacteria / fungus that, when administered in sufficient amounts as live bacteria to a host, can have a beneficial effect on the health of the host.

[0023] In certain embodiments, the additional probiotic bacteria is selected from Lactobacillus, Lactobacillus casei, Bifidobacterium, Lactobacillus mucosus, Lactobacillus plantarum, Lactobacillus unisomeris, Lactobacillus spp., Streptococcus, Lactococcus, Propionibacterium, Propionibacterium, Leuconostoc, Pediococcus, Weizmannella, Zoococcus, Staphylococcus, Bacillus, Acetobacter, Komagata, Gluconacetobacter, Gluconobacter, or any combination thereof.

[0024] In certain embodiments, the yeast is selected from Brettanomyces heterophylla ( Brettanomyces anomalus )、Saccharomyces cerevisiae( Saccharomyces cerevisiae ), Brettanomyces brusselsii ( Brettanomyces Brussels ), Candida asteroidea ( Starry white ), Schizosaccharomyces pombe ( Schizosaccharomyces pigeone), Zygosaccharomyces bayerischen Zygosaccharomyces bailii ), or any combination thereof.

[0025] In certain embodiments, the composition further comprises additional additives.

[0026] Those skilled in the art can select and adjust the additional additives according to the needs. In certain embodiments, the additional additives are nutrients.

[0027] In certain embodiments, the additional additive is capable of having a beneficial effect on the health of the host to which it is administered.

[0028] In certain embodiments, the additional additive is a nutrient selected from dietary fiber, prebiotics, protein, lipids, minerals, vitamins, plant extracts, or any combination thereof.

[0029] In certain embodiments, the mineral is selected from iron, zinc, potassium, sodium, calcium, magnesium, and any combination thereof.

[0030] In certain embodiments, the vitamin is selected from vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin A, vitamin C, vitamin D, vitamin E, vitamin K, and any combination thereof.

[0031] In certain embodiments, the composition is used as a fermentation agent (e.g., a fermentation agent for plant fermented products, a fermentation agent for dairy products). In such embodiments, the Weizmannella coagulans in the composition participates in the fermentation process as a fermentation agent. For example, in the process of preparing yogurt, the Weizmannella coagulans is fermented together with fresh milk as a probiotic to prepare yogurt.

[0032] In certain embodiments, the composition contains 10 2 Up to 10 20 The amount of CFU / dose present (e.g. 10 6 Up to 10 14 CFU / dose).

[0033] In certain embodiments, the bacteria of the genus Lactobacillus are selected from the group consisting of: Lactobacillus paracasei ( Lactobacillus cheese ), Lactobacillus acidophilus ( Lactobacillus acidophilus ), Lactobacillus brevis ( Lactobacillus short ), Lactobacillus jensenii ( Lactobacillus jensenii ), Lactobacillus iners ( Lactobacillus inert ), Lactobacillus casei( Lactobacillus casei ), Lactobacillus crispatus ( Lactobacillus crispatus ), Lactobacillus curvus ( Lactobacillus curvatus ), Lactobacillus delbrueckii ( Lactobacillus delbrueckii ), Lactobacillus fermentum ( Lactobacillus fermentum ), Lactobacillus gasseri ( Lactobacillus gasseri ), Lactobacillus helveticus ( Lactobacillus helveticus ), Lactobacillus johnsonii ( Lactobacillus johnsonii ), Lactobacillus plantarum ( Lactobacillus plantarum ), Lactobacillus reuteri ( Lactobacillus reuteri ), Lactobacillus rhamnosus ( Lactobacillus rhamnosus ), Lactobacillus sakei( Lactobacillus sakei ), Lactobacillus salivarius ( Lactobacillus salivarius ), or any combination thereof.

[0034] In certain embodiments, the bacterium of the genus Bifidobacterium is selected from the group consisting of: Bifidobacterium animalis ( Bifidobacteriumanimalium ), Bifidobacterium bifidum ( Bifidobacterium bifidum ), Bifidobacterium breve ( Bifidobacteriumbreve ), Bifidobacterium infantis ( Bifidobacterium infantis ), Bifidobacterium longum ( Bifidobacterium longum ), Bifidobacterium adolescentis ( Bifidobacterium adolescentis ), or any combination thereof.

[0035] In certain embodiments, the bacterium of the genus Bacillus is selected from: Bacillus subtilis ( Bacillus subtle ), or any combination thereof.

[0036] In certain embodiments, the bacteria of the genus Propionibacterium are selected from: Propionibacterium schizophyllae ( Propionibacteriumshermanii ), Propionibacterium freudenreichii ( Propionibacterium freudenreichii ), Propionibacterium acidipropionici ( Propionibacterium acidipropionici ), or any combination thereof.

[0037] In certain embodiments, the bacteria of the genus Streptococcus are selected from: Streptococcus thermophilus ( Streptococcus thermophilic ), Streptococcus salivarius ( Streptococcus salivarius ), or any combination thereof.

[0038] In certain embodiments, the bacterium of the genus Lactococcus is Lactococcus lactis ( Lactococcus lactis ).

[0039] In certain embodiments, the bacteria of the genus Enterococcus are selected from: Enterococcus faecalis ( Enterococcus faecalis ), Enterococcus faecium ( Enterococcus faecium ), or any combination thereof.

[0040] In a third aspect, the present application provides a food, comprising the aforementioned Weizmannella coagulans or the aforementioned composition.

[0041] In the text, the term "food" is used in a broad sense, including food and drink for humans, and also covers food and drink (ie feed) for animals. In certain embodiments, the food is suitable for and designed for human consumption.

[0042] It is to be understood that, depending on the purpose, application mode or administration mode, the food product of the present application can be in the form of liquid, solid or semi-solid.

[0043] Therefore, in certain embodiments, the food is a solid food (eg, soft candy, lozenge, capsule, bacterial powder), a liquid food (eg, a drink), or a semi-solid food (eg, jelly).

[0044] In certain embodiments, the food product is a dietary supplement, a nutraceutical, a functional food, or a beverage product.

[0045] In certain embodiments, the food is a dairy product (eg, yogurt, flavored fermented milk, lactic acid bacteria beverage, cheese).

[0046] In certain embodiments, the food product is formulated for oral administration.

[0047] In certain embodiments, the food product is in the form of a pill, powder, capsule, tablet, granule, film-coated tablet, sachet, or dragee.

[0048] In certain embodiments, the food further comprises prebiotics. Generally speaking, prebiotics are non-digestible, and they cannot be broken down and absorbed in the stomach or small intestine, so they can remain intact when reaching the colon through the stomach and small intestine. The example of prebiotics includes some oligosaccharides, such as oligofructose (FOS), inulin, oligoxylose (XOS), polydextrose or any mixture thereof.

[0049] In certain embodiments, the food may also include (but not limited to) one or any combination of the following substances: probiotics (e.g., probiotic bacteria), dietary fiber, protein (e.g., enzymes), carbohydrates, lipids (e.g., fats), vitamins, minerals, plant ingredients (e.g., plant extracts), amino acids, immunomodulators, milk substitutes, or metabolites or extracts of Weizmannella coagulans.

[0050] In certain embodiments, foods for use with the present invention may comprise a protein source such as animal protein (eg, milk, meat, or egg protein), plant protein (eg, soy, wheat, rice, or pea protein); a mixture of free amino acids; or a combination thereof.

[0051] In certain embodiments, food for the present invention may comprise a fat source, and the lipid constituting the fat source may be any suitable fat or fat mixture. Vegetable fats such as soybean oil, palm oil, coconut oil, safflower oil, sunflower oil, corn oil, canola oil and lecithin. If desired, animal fats such as milk fat may also be added.

[0052] In certain embodiments, food products for use with the present invention may include carbohydrates such as sucrose, lactose, glucose, fructose, corn syrup solids, maltodextrin, and mixtures thereof.

[0053] In certain embodiments, the food for use in the present invention may include dietary fiber. The dietary fiber may come from any suitable source, including, for example, soy, pea, oat, pectin, guar gum, gum arabic, oligofructose, oligogalactose, sialyllactose, and oligosaccharides derived from animal milk.

[0054] In certain embodiments, the food used in the present invention may also contain suitable vitamins and minerals, which may be included in the food in appropriate amounts.

[0055] In certain embodiments, the coagulant Weizmannella of the present invention can also be combined with various sweeteners or flavoring agents, coloring substances, stabilizers, glidants, fillers and other excipients acceptable in food.

[0056] In certain embodiments, the Weizmannella coagulans is present in the form of a concentrate.

[0057] In certain embodiments, the coagulated Weizmannella in the food is 10 2 Up to 10 20 The amount of CFU / dose present (e.g. 10 6 Up to 10 14 CFU / dose).

[0058] In a fourth aspect, the present application provides a pharmaceutical composition comprising the Weizmannella coagulans as described above or the composition as described above.

[0059] As used herein, the term "pharmaceutical" encompasses pharmaceuticals for use in humans as well as pharmaceuticals for use in animals (ie, veterinary applications). In certain embodiments, the pharmaceutical is for use in humans.

[0060] In certain embodiments, the pharmaceutical composition comprises a preparation of coagulated Weizmannella or a composition.

[0061] In certain embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable carrier.

[0062] In certain embodiments, the pharmaceutical composition is formulated for oral administration.

[0063] In certain embodiments, the pharmaceutical composition is in the form of a pill, powder, capsule, tablet, granule, film-coated agent, cream, ointment, gel, lotion, foam, suppository, sachet, or dragee.

[0064] In certain embodiments, the coagulated Weizmannella in the food is 10 2 Up to 10 20 The amount of CFU / dose present (e.g. 10 6 Up to 10 14 CFU / dose).

[0065] In a fifth aspect, the present application provides a culture comprising the Weizmannella coagulans as described above or the composition as described above.

[0066] In certain embodiments, the culture is a suspension of Weizmannella coagulans.

[0067] In certain embodiments, the culture further comprises components that provide nutrients (eg, solid or liquid culture medium, feeder cell layers).

[0068] In certain embodiments, the nutrient-providing ingredient is selected from protein, carbon source, nitrogen source, fat, vitamin, mineral, or any combination thereof.

[0069] In certain embodiments, the carbon source is selected from D-glucose, D-fructose, D-mannose, mannitol, D-cellobiose, or any combination thereof.

[0070] In certain embodiments, the nitrogen source is selected from diammonium phosphate, beef meal, tryptone, yeast powder, or any combination thereof.

[0071] In certain embodiments, the culture further comprises a cell-free culture filtrate that coagulates Weizmannella.

[0072] In certain embodiments, the culture further comprises a derivative of Weizmannella coagulans.

[0073] In certain embodiments, the derivative is selected from a metabolite, an enzyme, a cell structural component (e.g., a cell wall or a component thereof), an exopolysaccharide, a bacteriocin, a compound containing an immunogenic component, or any combination thereof.

[0074] In the sixth aspect, the present application provides the use of the aforementioned Weizmannella coagulans or the aforementioned composition or the aforementioned food or the aforementioned pharmaceutical composition or the aforementioned culture in the preparation of a drug, wherein the drug can inhibit pathogenic bacteria infection, or prevent and / or treat diseases and / or symptoms caused by pathogenic bacteria infection.

[0075] In another aspect, the present invention provides a method for inhibiting pathogen infection, or preventing and / or treating diseases and / or symptoms caused by pathogen infection, the method comprising administering an effective amount of the Weizmannella coagulans as described above, or the composition as described above, or the food as described above, or the pharmaceutical composition as described above, or the culture as described above to a subject in need.

[0076] In certain embodiments, the pathogen is a pathogenic bacteria.

[0077] In certain embodiments, the pathogenic bacteria are selected from the genus Escherichia coli (e.g., Escherichia coli ( Escherichia coli )), Staphylococci (e.g., Staphylococcus aureus ( Staphylococcus aureus ), Streptococcus spp., or any combination thereof.

[0078] In certain embodiments, the pathogen is a pathogenic fungus.

[0079] In certain embodiments, the pathogenic fungus is selected from yeast (e.g., Debaryomyces hansenii ( Debaryomyces hansenii ), Aspergillus spp., Epidermophyton spp., or any combination thereof.

[0080] In certain embodiments, the disease is selected from respiratory diseases (e.g., pneumonia, bronchitis), digestive diseases (e.g., enteritis, gastritis, gastric ulcer, Crohn's disease), internal organ infections (e.g., meningitis, pericarditis), skin and soft tissue infections (e.g., folliculitis, furuncle, carbuncle), or any combination thereof.

[0081] In a seventh aspect, the present invention provides a use of the aforementioned Weizmannella coagulans or the aforementioned composition in preparing a starter.

[0082] In certain embodiments, the Weizmannella coagulans as described above or the composition as described above is used as a starter culture for the food product.

[0083] In certain embodiments, the food is a solid food (eg, cheese, bread) or a liquid food (eg, yogurt, flavored fermented milk, lactic acid bacteria beverage).

[0084] Definition of terms

[0085] In the present invention, unless otherwise specified, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. Meanwhile, in order to better understand the present invention, the definitions and explanations of the relevant terms are provided below.

[0086] As used herein, the term "Weizmannella coagulans" refers to a lactic acid-producing bacterium known by the Latin name Weizmannia coagulans, which belongs to the genus Bacillus in taxonomy. Weizmannella coagulans is also known as "Bacillus coagulans ( Bacillus coagulans )"or" Heyndrickxia coagulans Therefore, in this paper, Weizmannia coagulans or Bacillus coagulans or Heyndrickxia coagulans All refer to the same strain, namely, Weizmannella coagulans. Generally, the cells of Weizmannella coagulans are rod-shaped, have terminal spores, no flagella, and are Gram-positive bacteria. They can decompose sugars to produce L-lactic acid and are homolactic fermentative bacteria.

[0087] As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier that is pharmacologically and / or physiologically compatible with a subject and an active ingredient, which is well known in the art (see, e.g., Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995), and includes, but is not limited to, pH regulators, surfactants, adjuvants, and ionic strength enhancers. For example, pH regulators include, but are not limited to, phosphate buffers; surfactants include, but are not limited to, cationic, anionic or nonionic surfactants, such as Tween-80; ionic strength enhancers include, but are not limited to, sodium chloride.

[0088] As used herein, the term "dietary supplement" refers to an edible product that can provide a beneficial effect (e.g., nutritional effect, preventive effect, therapeutic effect or other beneficial effect) to a consumer. In this article, dietary supplements include products such as nutritional products and supplements.

[0089] As used herein, the term "drug" covers drugs for both human and animal use in human and veterinary medicine, and also covers drugs for incorporation into animal feed (e.g., livestock feed and / or pet food). In addition, the term "drug" as used herein means any substance that provides a therapeutic, preventive, and / or beneficial effect. The term "drug" as used herein is not necessarily limited to substances that require a marketing approval, but includes substances that can be used in cosmetics, health products, foods (including, for example, feed and beverages), probiotic cultures, and dietary supplements.

[0090] As used herein, the term "CFU (Colony-Forming Units)" refers to the total number of microorganisms such as bacteria, fungi, yeast, etc. in a product, and is usually used for calculation of viable bacteria count.

[0091] As used herein, the term "CFU / dose" means the amount of bacteria present in a composition / food / pharmaceutical composition provided to a subject each day or each time. For example, in certain embodiments, the amount of coagulant Weizmannella in the food is 10 6 Up to 10 14 The amount of CFU / dose present (e.g. 10 7 Up to 10 10 In this embodiment, if the Weizmannella coagulans is administered in a food (e.g., a solid beverage, yogurt), the food (e.g., a solid beverage, yogurt) provided to the subject every day or each time may contain about 10 CFU / dose. 6 Up to 10 14 Of course, alternatively, this amount of bacteria can be divided into multiple administrations, as long as the total amount of coagulant Weizmannella received by the subject in any particular time (e.g., every 24-hour period) is from about 10 6 Up to 10 14 CFU of bacteria, i.e., Weizmannella coagulans in food products or dietary supplements meeting the above requirements, is 10 6 Up to 10 14 The amount of CFU / dose present (e.g. 10 7 Up to 10 10 CFU / dose). Advantageous Effects of the Invention

[0092] Compared with the coagulant Weizmannella in the prior art, the coagulant Weizmannella of the present application has better tolerance to the acidic environment of gastric juice and bile salts, and also has better heat resistance, so it can adapt to the gastrointestinal environment and play a colonization role. In addition, the coagulant Weizmannella is sensitive to antibiotics and does not have toxin genes, so the coagulant Weizmannella has high food safety and has great application potential in the food field. Furthermore, the coagulant Weizmannella also has a certain inhibitory effect on pathogenic bacteria and fungi, so the coagulant Weizmannella has great prevention and / or treatment potential in diseases caused by pathogenic bacteria infection.

[0093] Embodiments of the present invention will be described in detail below in conjunction with the accompanying drawings and examples, but it will be appreciated by those skilled in the art that the following drawings and examples are only used to illustrate the present invention, rather than to limit the scope of the present invention. Various objects and advantages of the present invention will become apparent to those skilled in the art based on the following detailed description of the accompanying drawings and preferred embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0094] Figure 1 Phylogenetic analysis of 16S rRNA of Weizmannella coagulans We06.

[0095] Figure 2 This is the colony morphology of Weizmannella coagulans We06.

[0096] Figure 3 The results of antibiotic sensitivity analysis of Weizmannella coagulans We06.

[0097] Figure 4 The results of whole genome analysis of Weizmannella coagulans We06.

[0098] Figure 5 To condense the survival rate of Weizmannella We06 under different temperature treatments when using Anmuxi as the matrix.

[0099] Figure 6 To condense the survival rate of Weizmannella We06 under different temperature treatments when sucrose was used as the matrix.

[0100] Figure 7 This is the result of the acid tolerance analysis of Weizmannella We06.

[0101] Figure 8 This is the result of the analysis of the tolerance of Weizmannella We06 to bile salts.

[0102] Fig. 9 The results show the inhibitory effect of Weizmannella coagulans We06 on Escherichia coli, Staphylococcus aureus and Debaryomyces hansenii.

[0103] Sequence information

[0104] The information of the partial sequences involved in the present invention is provided in Table 1 below.

[0105] Table 1: Description of sequences

[0106] Notes on the Deposit of Biological Materials

[0107] Weizmannella coagulans We06 ( Weizmannia coagulans We06) has been deposited at the China General Microbiological Culture Collection Center (CGMCC) located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. It has the deposit number CGMCC No.28819 and the deposit date is October 30, 2023. DETAILED DESCRIPTION

[0108] The invention will now be described with reference to the following examples which are intended to illustrate the invention rather than to limit the invention.

[0109] Unless otherwise indicated, the experiments and procedures described in the examples were performed essentially according to conventional methods well known in the art and described in various references. For example, conventional techniques of immunology, biochemistry, chemistry, molecular biology, microbiology, cell biology, genomics and recombinant DNA used in the present invention can be found in Sambrook, Fritsch and Maniatis, MOLECULAR CLONING: A LABORATORY MANUAL, 2nd edition (1989); CURRENT PROTOCOLS IN MOLECULAR BIOLOGY (FM Ausubel et al., eds., (1987)); METHODS IN ENZYMOLOGY series (Academic Press): PCR 2: A PRACTICAL APPROACH (MJ MacPherson, BD Hames and GR Taylor, eds. (1995)); and ANIMAL CELL CULTURE (Animal Cell Culture). CELLCULTURE) (RI Freshney, ed. (1987)). Or, for example, conventional techniques such as biochemistry, microbiology, proteomics and genomics used in the present invention can be found in JF Robyt and BJ White, Biochemical Techniques: Theory and Practice, 2nd edition (1990); MT Madigan, JM Martinko, KS Bender, DH Buckley, and DA Stahl, Brock Biology of Microorganisms, 15th edition (2017); GJ Pelczar, ECSChan and NR Krieg, Laboratory Experiments in Microbiology, 5th edition (1993); SR Pennington and MJ Dunn, Proteomics: From Protein Sequence to Function, 2001, etc.

[0110] In addition, if the specific conditions are not specified in the examples, they are carried out according to the conventional conditions or the conditions recommended by the manufacturer. If the manufacturer is not specified in the reagents or instruments used, they are all conventional products that can be obtained commercially. It is known to those skilled in the art that the embodiments describe the present invention by way of example and are not intended to limit the scope of the present invention. All public cases and other references mentioned herein are incorporated herein by reference in their entirety.

[0111] Example 1. Phenotypic and Identification Analysis of Weizmannella coagulans

[0112] Lactic acid bacteria We06 was isolated from farm dairy products in Xinjiang. A single clone was picked from the plate culture medium of lactic acid bacteria We06 and transferred to 5 mL of liquid MRS medium (peptone 10 g / L (Bioengineering Co., Ltd.), beef powder 5 g / L (Beijing Aoboxing Biotechnology Co., Ltd.), glucose 20 g / L, anhydrous sodium acetate 5 g / L, manganese sulfate 0.05 g / L, potassium dihydrogen phosphate 2 g / L, ammonium citrate 2 g / L, Tween 80 1mL (Tianjin Komiou Chemical Reagent Co., Ltd.), yeast powder 4 g / L (Thermo Fisher Scientific), magnesium sulfate 0.2 g / L (Tianjin Tianli Chemical Reagent Co., Ltd.), pH adjusted to 6.2-6.4), and cultured in a constant temperature shaker at 43°C for 14 h. 1.5 mL of bacterial solution was aspirated from the shaking tube and centrifuged at 10,000 rpm for 3 min, and the supernatant was discarded, leaving the bacterial precipitate. The MP bacterial genomic DNA extraction kit was used to extract genomic DNA, and the bacterial universal primers 27F (SEQ ID NO: 2) and 1495R (SEQ ID NO: 3) were used to amplify the bacterial 16S rRNA. The DNA was sent to a biological company for 16S rRNA sequencing. The obtained 16S rRNA sequence is shown in SEQ ID NO: 1.

[0113] After NCBI Blast alignment of the 16S rRNA sequence of lactic acid bacteria We06, the sequences of five strains with high homology to the NCBI alignment results and strains of different families and genera were selected to construct a phylogenetic tree using MEGA ( Figure 1 ). The results of the phylogenetic tree showed that lactic acid bacteria We06 and Weizmannia coagulans (Former name: Heyndrickxia coagulans ) were clustered on the same branch on the evolutionary tree, so the lactic acid bacteria We06 were identified as Weizmannella coagulans We06.

[0114] The Weizmannella We06 monoclonal was picked up with an inoculation loop and drawn on MRS solid medium (peptone 10 g / L (Bioengineering Co., Ltd.), beef powder 5 g / L (Beijing Aoboxing Biotechnology Co., Ltd.), glucose 20 g / L, anhydrous sodium acetate 5 g / L, manganese sulfate 0.05 g / L, potassium dihydrogen phosphate 2 g / L, ammonium citrate 2 g / L, Tween 801mL (Tianjin Komiou Chemical Reagent Co., Ltd.), yeast powder 4 g / L (Thermo Fisher Scientific), magnesium sulfate 0.2 g / L (Tianjin Tianli Chemical Reagent Co., Ltd.), agar powder 15 g / L (Beijing Lanjieke Technology Co., Ltd.)) by the line drawing method, and then placed in a 43°C incubator for 5 days to observe its phenotype. Its appearance is white and opaque, with irregular edges and rough protrusions on the surface. The diameter of the bacteria can reach 3.97 mm ( Figure 2 ).

[0115] Example 2. Analysis of carbon and nitrogen source utilization by Weizmannella coagulans We06

[0116] The API 50CHL kit (Mérieux, France, catalog number: REF CN5041010) was used to compare the carbohydrate metabolism of Weizmannella coagulans We06 using 49 biochemical tests. The strain was collected from the solid culture medium using a cotton swab and added to the suspension (2 mL) to prepare a high-concentration bacterial suspension (S), and the high-concentration bacterial suspension (S) was added to the suspension (5 mL) to prepare a bacterial suspension with a turbidity equivalent to 2McF, and the number of drops n was recorded. The 50CHL culture medium ampoule was opened and 2n drops of the above bacterial solution were added for inoculation. The bacterial suspension was added to the test wells on the test strip using a sterile pipette, sealed with paraffin, and placed in a 43°C incubator for culture. The reaction was observed at 24 h and 48 h, respectively. The main carbon sources used by We06 are D-glucose, D-fructose, D-mannose, mannitol, and D-cellobiose (Table 2).

[0117] Table 2 Analysis of the utilization of 49 carbon sources by Weizmannella coagulans We06

[0118] Note: '++': excellent growth; '+': growth; '+(w)': difficult growth; '-': no ​​growth.

[0119] Furthermore, four organic nitrogen sources, urea, beef powder, tryptone, and yeast powder, and three inorganic nitrogen sources, ammonium chloride, diammonium phosphate, and ammonium sulfate, were selected. By controlling the sole nitrogen source to configure different culture media, the logarithmic phase Weizmannella coagulans We06 was transferred to the culture medium containing the sole nitrogen source at a ratio of 1:100, placed at 43°C, 170 rpm shaking culture for 24 hours, and its absorbance was measured. Weizmannella coagulans We06 can better utilize diammonium phosphate, beef powder, tryptone, and yeast powder, but does not utilize ammonium sulfate, ammonium chloride, and urea, and the nitrogen source utilization is similar (Table 3).

[0120] Table 3 Nitrogen source analysis of Weizmannella coagulans We06

[0121] Note: '-': no ​​nitrogen source used; '+': available nitrogen source; '++': easily available nitrogen source; '+++': very easily available nitrogen source.

[0122] Example 3. Analysis of the sensitivity of Weizmannella coagulans We06 to antibiotics

[0123] The investigation of antibiotic sensitivity is an important safety aspect of bacteria used in food. The sensitivity of Weizmannella coagulans We06 to six antibiotics, chloramphenicol, ampicillin, erythromycin, penicillin G, tetracycline, and streptomycin, was tested using drug sensitivity test strips. A single clone of Weizmannella coagulans We06 was selected and placed in MRS liquid culture medium, cultured at 43°C and 170 rpm for 10 hours, the concentration of the diluted bacteria was adjusted, and it was spread on MRS solid culture medium to cover the entire plate surface. The drug sensitivity test strips were placed on the plates with tweezers in turn, and cultured and observed.

[0124] By measuring the diameter of the inhibition zone, it was found that Weizmannella coagulans We06 was sensitive to six antibiotics (chloramphenicol, ampicillin, erythromycin, penicillin G, tetracycline, and streptomycin). Figure 3 , Table 4). This means that the safety of Weizmannella coagulans We06 is relatively high and suitable for adding to food.

[0125] Table 4 Diameter of the inhibition zone of Weizmannella flocculata We06 (mm) and sensitivity of antibiotics

[0126] Example 4. Analysis of the genome, drug resistance genes and virulence genes of Weizmannella coagulans We06

[0127] The genome of Weizmannella coagulans We06 was extracted using a bacterial genome extraction kit and sent for testing. The software fastp (https: / / github.com / OpenGene / fastp) was used to perform quality control on the sequencing sequences at both ends, the software spades (https: / / github.com / ablab / spades) was used to splice the quality-controlled files, and Prokka (https: / / github.com / tseemann / prokka) was used to perform functional annotation on the genome of Weizmannella coagulans We06. MASH was used to compare the strain We06 with 157,652 assembled gene sequences of GTDB species classification, and the genome information with a mash distance less than 0.03 was obtained. The species classification information was determined by the Least Common Ancestors (LCA) analysis. The genome circle map of Weizmannella coagulans We06 was displayed using CG View (https: / / cgview.ca), and the drug-resistant gene database (https: / / ardb.cbcb.umd.edu) was used to compare the genes of Weizmannella coagulans We06 to mine the drug-resistant genes of Weizmannella coagulans We06. The genes of Weizmannella coagulans We06 were compared with the virulence factor database (http: / / www.mgc.ac.cn / VFs) to mine the virulence factors of Weizmannella coagulans We06.

[0128] The genome of Weizmannella coagulans We06 is 3716246 bp long, with a GC content of 46.26%, 84 tRNAs, 30 rRNAs, and 3541 CDSs. It was identified by MASH species as Weizmannia coagulans (Also known as: Heyndrickxia coagulans ) ( Figure 4 After the assembled genome data were imported into the CARD database (https: / / card.mcmaster.ca / ) for analysis and prediction, only four drug-resistant genes were found, with coverage higher than 99.00%, but similarities lower than 60.00% (Table 5), which showed low similarity with the genes in the database.

[0129] Table 5 Analysis of drug resistance genes of Weizmannella coagulans We06

[0130] The genome data were imported into the virulence factor database VFDB for analysis and prediction, and 7 virulence genes were predicted (Table 6), but there were no toxin genes such as bce T. ces , ent FM and cytK, indicating that the strain provided by the present invention has a low possibility of secreting toxins. When the strain is added to food, the strain does not secrete toxins and will not affect human health. In summary, Weizmannella coagulans We06 has food safety.

[0131] Table 6 Virulence gene analysis of Weizmannella coagulans We06

[0132] In summary, Weizmannella coagulans We06 does not have dangerous drug-resistant genes and toxin genes. These results indicate that the isolated Weizmannella coagulans We06 has potential probiotic properties and can be used as a candidate probiotic for probiotic drinks.

[0133] Example 5. Analysis of the heat resistance of Weizmannella coagulans We06 spores

[0134] Weizmannella coagulans We06 monoclone was selected and placed in MRS liquid medium, cultured at 43℃, 170 rpm for 10 h. The inoculation amount was transferred to the spore-producing medium at 12%, cultured at 43℃, 170 rmp for 48 h, and treated in an 80℃ metal bath for 15 min, which was the Weizmannella coagulans spore group. Amul yogurt and sucrose solution (30%, pH=3.4) were used as the matrix, and the survival rate after heat treatment at 78℃ for 40 min, 80℃ for 30 min, and 95℃ for 5 min was measured. The spores of Weizmannella coagulans We06 were heat-treated, and the untreated spores were used as blank controls. The survival rate was calculated by coating on MRS plates.

[0135] When using Amul yogurt (pasteurized heat-treated flavored yogurt) as the matrix, the survival rate of Weizmannella coagulans We06 was 84.56% after treatment at 78℃ for 40 min. The survival rates of Weizmannella coagulans We06 were 78.93% and 70.47% under the treatment conditions of 80℃ for 30 min and 95℃ for 5 min, respectively (Figure 5). Therefore, the spores of Weizmannella coagulans We06 have a high heat resistance in Amul yogurt and have a good survival rate under several temperature conditions.

[0136] When sucrose solution (30%, pH=3.4) was used as the matrix, the survival rate of Weizmannella coagulans We06 was 71.88% after treatment at 78℃ for 40min. After treatment at 80℃ for 30min, the survival rate of Weizmannella coagulans We06 was 83.16% (Figure 6). Therefore, Weizmannella coagulans We06 showed good heat resistance under 80℃ for 30min heat treatment.

[0137] Example 6. Analysis of the heat-resistant gene of Weizmannella coagulans We06

[0138] By performing high-throughput sequencing (including second-generation and third-generation sequencing technologies) and genome assembly on Weizmannella coagulans We06, and using Prokka software to predict genes and annotate their functions, we found that its heat resistance mechanism mainly consists of six aspects (Table 7). ikB and yrA , the encoded DNA gyrase introduces negative supercoils during replication and transcription to maintain DNA stability and prevent DNA denaturation or breakage at high temperatures, thereby ensuring genome integrity. Secondly, through the overexpression of cold shock proteins, key genes cspA It plays a major role in responding to low temperature stress, but it can also help maintain the homeostasis of proteins and nucleic acids under high temperature stress, and help bacteria adapt to the environment and survive under high temperature conditions by participating in the universal stress response pathway. In addition, phosphate metabolism has a significant impact on the survival ability of bacteria in high temperature environments. phoP, phoR, phoH2, phoH, phoA By sensing and responding to changes in phosphate concentration in the environment, the expression of related genes is regulated, thereby affecting the stress response and adaptability of bacteria. In addition, maintaining protein structural stability is another important aspect. Key genes clpC, yclP, clpP, clpB, clpE, clpX The encoded Clp protein family plays a dual role as a molecular chaperone and protease in cells, improving the survival rate of bacteria in high temperature environments by degrading misfolded or damaged proteins and helping new proteins to fold correctly. When cells are subjected to high temperature stress, heat shock proteins protect cells from heat damage by helping proteins to fold correctly, preventing protein aggregation, and repairing damaged proteins. Key genes include dnaJ, dnaK, hslO, mcsA, mcsB, rpoE, rpoD, rpoN, groEL, groES These genes play a key role in the high temperature stress response and improve the resistance of bacteria to heat stress. Finally, antioxidant and stress proteins are expressed through key genes opuC, opuBD, opuA, mutM, uspA, katE The encoded antioxidant enzymes and general stress proteins scavenge reactive oxygen species (ROS) and respond to environmental stress under high temperature conditions, protecting cells from oxidative damage and other environmental stresses and improving bacterial survival.

[0139] Table 7 Thermoresistant genes of Weizmannella coagulans We06

[0140] In summary, Weizmannella coagulans We06 can effectively survive in high temperature environments by maintaining the structural stability of DNA and proteins, regulating phosphate metabolism, and expressing cold shock proteins, heat shock proteins, and antioxidant and stress proteins. These findings not only reveal the complexity of its heat resistance mechanism, but also provide important basic data for further research and application.

[0141] Example 7. Analysis of acid resistance of Weizmannella coagulans We06

[0142] Pick a single colony of Weizmannella We06 and transfer it to 3 mL of MRS liquid medium. Culture it at 43°C overnight for 10 h. Cultivate it at 43°C until the OD 600 =1.2~1.4, transfer the bacterial liquid to the spore-forming medium at an inoculum volume of 12%, and culture at 43℃ for 48h; in simulated gastric juice (1 mol / L hydrochloric acid, pepsin, to a concentration of 10 g / L. pH=2.0, 3.0, 4.0 (NaOH adjustment), sterilized with a filter membrane (0.22 μm))) and incubate at 37℃ for 1, 2, and 3h, then spread and count, and repeat three times. The survival rate (SR) was calculated according to the following formula: SR=(Nt / N0)×100%, where Nt (CFU / mL) is the number of viable bacteria after treatment for t hours, and N0 (CFU / mL) is the number of viable bacteria before treatment.

[0143] After 3 h of shaking culture in simulated gastric fluid with different acidity, the results were as follows: Figure 7 As shown. Under the extreme condition of pH=2, the survival rate of Weizmannella coagulans We06 was still 25.11% (Figure 7). At pH=3, the survival rate of Weizmannella coagulans We06 was 51.98%, and the survival rate of Weizmannella coagulans We06 in an environment of pH 3 was better than that under the condition of pH=2. At pH=4, the survival rate of Weizmannella coagulans We06 was 78.41%. It can be seen that in the simulated gastric juice environment of different acidity, Weizmannella coagulans We06 showed good acid resistance.

[0144] Example 8. Bile salt tolerance of Weizmannella coagulans We06

[0145] A single clone of Weizmannella coagulans We06 was selected and placed in MRS liquid medium and cultured overnight at 43°C for 10 h. The overnight cultured bacterial solution was transferred to the spore-forming medium at a 12% inoculation rate and cultured at 43°C and 170 rpm for 48 h. 100 μL of the bacterial solution was then placed in 900 μL of aqueous solutions with different bile salt concentrations and placed at 45°C. The mixture was shaken and mixed at 0 h, 1 h, 2 h, and 3 h, and 100 μL of the sample was taken and incubated with ddH 2 O gradient dilution (10, 10 2 , 10 3 , 104 , 10 5 ), take the diluted sample (10 μL) for spot counting, and use 0% bile salt concentration as the control; count and calculate the survival rate.

[0146] Survival rate = number of viable bacteria in the culture solution treated for 3 hours at different bile salt concentrations / number of viable bacteria in the culture solution treated for 3 hours at 0% bile salt concentration * 100%

[0147] The survival rate of Weizmannella coagulans We06 in different bile salt contents showed that in 0.3% bile salt, the survival rate of Weizmannella coagulans We06 was 40% ( Figure 8 ). In 0.5% bile salt, the survival rate of Weizmannella coagulans We06 was 51.28%; in 1.0% bile salt, the survival rate of Weizmannella coagulans We06 was 15.38%. Therefore, Weizmannella coagulans We06 has a certain tolerance to bile salt.

[0148] Example 9. Analysis of the antibacterial effect of coagulating Weizmannella We06

[0149] After activating Weizmannella coagulans We06, use a ruler and a marker to draw two straight lines with a length of 2 cm and a spacing of 1.5 cm on the MRS plate. Then pick out the activated single colony of Weizmannella coagulans We06 and draw lines along the drawn straight lines.

[0150] Indicator bacteria (Staphylococcus aureus) were cultured in LB liquid medium. Staphylococcus aureus (donated from the Institute of Microbiology, Chinese Academy of Sciences), Escherichia coli Escherichiacoli (gift from the Institute of Microbiology, Chinese Academy of Sciences), Debaryomyces hansenii Debaryomyces hansenii (Purchased from China General Microbiological Culture Collection Center CGMCC No.23966)), cultured to OD 600 The ratio of LB to LB (0.7% agar) was about 0.6, and then Escherichia coli and Staphylococcus aureus were inoculated into the semisolid LB (0.7% agar) cooled to about 45°C at a ratio of 1:100, and Debaryomyces hansenii was inoculated into the semisolid PDA (0.7% agar) cooled to about 45°C at a ratio of 1:200, and the mixture was poured onto the upper plate. The double-layer plates containing Escherichia coli and Staphylococcus aureus were placed in a 37°C incubator for 13 h, and the double-layer plates containing Hansenii were placed in a 30°C incubator for 13 h. The diameter of the antibacterial circle close to the circle was recorded, and the approximate length and width of the irregular antibacterial circle were measured. Finally, the relative strength of the antibacterial effect was compared by area.

[0151] By calculating the inhibition area, it can be seen that the inhibition area of ​​Weizmannella coagulans We06 against Escherichia coli is 4159.32 mm 2 The inhibition area of ​​Staphylococcus aureus is 2284.22 mm2 , the inhibition area of ​​Debaryomyces hansenii is about 300mm 2 ( Fig. 9 ). Therefore, the coagulant Weizmannella We06 of the present application has good inhibitory activity against bacteria, and the inhibitory effect on Escherichia coli is the best, followed by Staphylococcus aureus; on this basis, the coagulant Weizmannella We06 of the present application also has certain inhibitory activity against fungi, for example, Debaryomyces hansenii.

[0152] Example 10. Analysis of sporulation genes of Weizmannella coagulans We06

[0153] First, the sample genome of Weizmannella coagulans We06 was extracted. The specific operation includes: centrifugation at 12,000 rpm for 2 min to collect cells in the logarithmic growth phase and discard the supernatant, adding 500 μL of Buffer BP to resuspend the cells, adding 50 μL of Lysozyme (20 mg / mL) and mixing thoroughly, and incubating in a 37°C water bath for 60 min, inverting and mixing every 10 min. Then centrifuge at 12,000 rpm for 5 min at room temperature and discard the supernatant. Then add 180 μL of Buffer LS-2, 20 μL of Proteinase K (20 mg / mL) and 10 μL of RNase A (10 mg / mL), oscillate or blow to mix thoroughly, and incubate in a 56°C water bath for 10 min until the solution becomes transparent. If the solution does not become transparent, continue to lyse for 30 min, oscillating or blowing to mix every 5 min. Then, add 200 μL of Buffer BS-2 and 200 μL of 100% ethanol to the solution, mix thoroughly, transfer the solution to the Bacterial DNA Mini Column, let stand at room temperature for 1 min, centrifuge at 12,000 rpm for 1 min and discard the filtrate. Add 500 μL of Buffer WA to the Mini Column, centrifuge at 12,000 rpm for 1 min, and discard the filtrate. Then add 750 μL of Buffer WB, centrifuge at 12,000 rpm for 1 min, discard the filtrate, and repeat this step once. Place the Mini Column on a new 2 mL Collection Tube and centrifuge at 12,000 rpm for 2 min to improve RNA purity. Then place the Mini Column on a new 1.5 mL centrifuge tube, add 50 μL of ElutionBuffer or sterile water to the center of the membrane, let stand at room temperature for 1 min, and centrifuge at 12,000 rpm for 2 min to elute the DNA. If a larger yield is required, the centrifuge solution can be added back to the center of the membrane, allowed to stand for 2 minutes, and then centrifuged again.

[0154] After extracting the genome of the Weizmannella coagulans sample, we performed second-generation error correction and third-generation splicing on its genome, and then used Prokka software for gene prediction and functional annotation (Table 8), revealing the complexity of its sporulation mechanism. The analysis showed that the mechanism mainly includes the following six aspects: the initiation phase consists of KinA, KinB, KinC and KinE The sensor kinases encoded by these genes initiate the sporulation process by sensing environmental signals and activating the phosphate signaling pathway. Spo0A, Spo0B and Spo0F Genes play a key role in signal transduction, including Spo0A It is an important transcriptional regulatory factor that regulates the expression of downstream genes after phosphorylation activation; the key genes in the early division stage are FWf , which participates in the formation of septa during cell division. SpoIIAA, SpoIIAB and SpoIIAC Regulation of gene-encoded proteins σF factor, σF It is a forespore-specific sigma factor responsible for the expression of forespore genes; during the differentiation of forespores and mother cells, SpoIIM, SpoIIP and SpoIID The gene promotes the formation of the forespore envelope, ensuring the interaction between the forespore and the mother cell. SpoIIIAE, SpoIIIAF, SpoIIIAH, SpoIIIAD, SpoIIIAB and SpoIIIAC Genes involved in mother cell to forespore signaling, ensuring correct gene expression; spore shell formation depends on SpoIVA Gene, which plays a key role in the initial formation of the spore coat, helping to build the outer structure of the spore. CotA, CotB, CotC, CotD, CotE, CotF, CotG, CotH, CotI, CotJA, CotJB, CotJC and CotY_Z The proteins encoded by the genes make up the spore coat, providing mechanical protection and environmental resistance; late differentiation and maturation stages involve SpoVAA, SpoVAB, SpoVAC, SpoVAD, SpoVAE and SvD Genes involved in the dehydration and mineralization of spores, enhancing the durability of spores. ikB Genetically encoded σH Factors play a key role in regulating the expression of many genes during late sporulation; finally, environmental sensing and stress responses are controlled by SigB Gene-mediated, the gene encoding σB Factors involved in cellular responses to environmental stress.

[0155] Table 8 Sporulation genes of Weizmannella coagulans We06

[0156] In summary, through these genes and the proteins they encode, Weizmannella coagulans can effectively sense environmental changes and respond accordingly, ensuring the success of its sporulation process and the viability of spores.

[0157] Although the specific embodiments of the present invention have been described in detail, it will be understood by those skilled in the art that various modifications and changes may be made to the details according to all the teachings that have been published, and these changes are within the scope of protection of the present invention. The entire invention is given by the attached claims and any equivalents thereof.

Claims

1. A coagulant Weizmannella Weizmannia coagulans ), the coagulant Weizmannella is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number being CGMCC No. 28819.

2. The Weizmannella coagulans of claim 1, having one or more characteristics selected from the following: (1) The colonies are white, with uneven edges and rough surfaces; (2) Tolerance to gastric acid and / or bile salts; (3) Heat resistance; (4) Having the ability to produce spores; (5) Ability to inhibit pathogenic bacteria and / or fungi; Preferably, the pathogenic bacteria is selected from Staphylococcus aureus ( Staphylococcus aureus ), Escherichia coli ( Escherichiacoli ), or any combination thereof; Preferably, the pathogenic fungus is Debaryomyces hansenii ( Debaryomyces hansenii ).

3. The Weizmannella coagulans according to claim 1 or 2, comprising: a) the nucleotide sequence shown in SEQ ID NO: 1; or, b) a nucleotide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, at least 99.95%, at least 99.99% or 100% identical to the nucleotide sequence of SEQ ID NO: 1; or, c) a complementary nucleotide sequence of a) or b).

4. A composition comprising the Weizmannella coagulans according to any one of claims 1 to 3; Preferably, the composition further comprises additional probiotic bacteria and / or probiotic fungi (such as yeast), wherein The additional probiotic bacteria and / or probiotic fungi are edible; Preferably, the additional probiotic bacteria are selected from the group consisting of Lactobacillus, Lactobacillus casei, Bifidobacterium, Lactobacillus mucis, Lactobacillus plantarum, Lactobacillus unisomeris, Lactobacillus spp., Streptococcus, Lactococcus, Propionibacterium, Propionibacterium, Leuconostoc, Pediococcus, Weizmannella, Zoococcus, Staphylococcus, Bacillus, Acetobacter, Komagata, Gluconacetobacter, Gluconobacter, or any combination thereof; Preferably, the bacteria of the genus Lactobacillus are selected from: Lactobacillus paracasei ( Lactobacillus paracasei ), Lactobacillus acidophilus ( Lactobacillus acidophilus ), Lactobacillus brevis ( Lactobacillus brevis ), Lactobacillus jensenii ( Lactobacillus jensenii ), Lactobacillus iners ( Lactobacillus iners )、Lactobacillus casei( Lactobacillus casei ), Lactobacillus crispatus ( Lactobacillus crispatus ), Lactobacillus curvus ( Lactobacillus curvatus ), Lactobacillus delbrueckii ( Lactobacillus delbrueckii ), Lactobacillus fermentum ( Lactobacillusfermentum ), Lactobacillus gasseri ( Lactobacillus gasseri ), Lactobacillus helveticus ( Lactobacillus helveticus ), Lactobacillus johnsonii ( Lactobacillus johnsonii ), Lactobacillus plantarum ( Lactobacillus plantarum ), Lactobacillus reuteri ( Lactobacillus reuteri ), Lactobacillus rhamnosus ( Lactobacillus rhamnosus ), Lactobacillus sakei( Lactobacillus sakei ), Lactobacillus salivarius ( Lactobacillus salivarius ), or any combination thereof; Preferably, the bacteria of the genus Bifidobacterium are selected from: Bifidobacterium animalis ( Bifidobacterium animalis ), Bifidobacterium bifidum ( Bifidobacterium bifidum ), Bifidobacterium breve ( Bifidobacterium breve ), Bifidobacterium infantis ( Bifidobacterium infantis ), Bifidobacterium longum ( Bifidobacterium longum ), Bifidobacterium adolescentis ( Bifidobacterium adolescentis ), or any combination thereof; Preferably, the bacterium of the genus Bacillus is selected from: Bacillus subtilis ( Bacillus subtilis ), or any combination thereof; Preferably, the bacteria of the genus Propionibacterium are selected from: Propionibacterium schizophyllae ( Propionibacterium shermanii ), Propionibacterium freudenreichii ( Propionibacterium freudenreichii ), Propionibacterium acidipropionici ( Propionibacterium acidipropionici ), or any combination thereof; Preferably, the bacteria of the genus Streptococcus are selected from: Streptococcus thermophilus ( Streptococcus thermophilus ), Streptococcus salivarius ( Streptococcus salivarius ), or any combination thereof; Preferably, the bacteria of the genus Lactococcus are Lactococcus lactis ( Lactococcus lactis ); Preferably, the yeast is selected from Brettanomyces heterophylla ( Brettanomyces anomalus )、Saccharomyces cerevisiae( Saccharomyces cerevisiae ), Brettanomyces brusselsii ( Brettanomyces bruxellensis ), Candida asteroidea ( Candida stellata ), Schizosaccharomyces pombe ( Schizosaccharomyces pomb e), Zygosaccharomyces bayerischen Zygosaccharomyces bailii ), or any combination thereof; Preferably, the composition further comprises additional additives; Preferably, the additional additive is a nutrient selected from dietary fiber, prebiotics, protein, lipids, minerals, vitamins, plant extracts, or any combination thereof.

5. A food comprising the coagulant Weizmannella according to any one of claims 1 to 3 or the composition according to claim 4; Preferably, the food is a solid food (e.g., soft candy, lozenge, capsule, bacterial powder), a liquid food (e.g., a drink), or a semi-solid food (e.g., jelly); Preferably, the food is a dietary supplement, a nutritional preparation, a functional food or a beverage product; Preferably, the food is a dairy product (e.g., yogurt, flavored fermented milk, lactic acid bacteria beverage, cheese); Preferably, the food product is formulated for oral administration; Preferably, the food is in the form of pills, powders, capsules, tablets, granules, film-coated tablets, sachets or dragees; Preferably, the food further comprises prebiotics; Preferably, the coagulated Weizmannella in the food is 10 2 Up to 10 20 The amount of CFU / dose present (e.g. 10 6 Up to 10 14 CFU / dose).

6. A pharmaceutical composition comprising the Weizmannella coagulans according to any one of claims 1 to 3 or the composition according to claim 4; Preferably, the pharmaceutical composition comprises a preparation of the coagulated Weizmannella or the composition; Preferably, the pharmaceutical composition comprises a pharmaceutically acceptable carrier; Preferably, the pharmaceutical composition is formulated for oral administration; Preferably, the pharmaceutical composition is in the form of pills, powders, capsules, tablets, granules, film-coated agents, creams, ointments, gels, lotions, foams, suppositories, sachets or dragees; Preferably, the coagulated Weizmannella in the food is 10 2 Up to 10 20 The amount of CFU / dose present (e.g. 10 6 Up to 10 14 CFU / dose).

7. A culture comprising the Weizmannella coagulans according to any one of claims 1 to 3 or the composition according to claim 4; Preferably, the culture is a bacterial suspension of Weizmannella coagulans; Preferably, the culture further comprises components that provide nutrients (e.g., solid or liquid culture medium, feeder cell layer); Preferably, the nutrient-providing ingredients are selected from proteins, carbon sources, nitrogen sources, fats, vitamins, minerals, or any combination thereof; Preferably, the carbon source is selected from D-glucose, D-fructose, D-mannose, mannitol, D-cellobiose, or any combination thereof; Preferably, the nitrogen source is selected from diammonium phosphate, beef powder, tryptone, yeast powder, or any combination thereof; Preferably, the culture further comprises a cell-free culture filtrate of coagulated Weizmannella; Preferably, the culture further comprises a derivative of Weizmannella coagulans; Preferably, the derivative is selected from a metabolite, an enzyme, a cell structural component (eg, a cell wall or a component thereof), an exopolysaccharide, a bacteriocin, a compound containing an immunogenic component, or any combination thereof.

8. Use of the Weizmannella coagulans according to any one of claims 1 to 3, the composition according to claim 4, the food according to claim 5, the pharmaceutical composition according to claim 6, or the culture according to claim 7 in the preparation of a medicament capable of inhibiting pathogenic bacteria infection, or preventing and / or treating diseases and / or symptoms caused by pathogenic bacteria infection; Preferably, the pathogen is a pathogenic bacterium; Preferably, the pathogenic bacteria are selected from the genus Escherichia coli (e.g., Escherichia coli ( Escherichiacoli )), Staphylococci (e.g., Staphylococcus aureus ( Staphylococcus aureus ), Streptococcus spp., or any combination thereof; Preferably, the pathogen is a pathogenic fungus; Preferably, the pathogenic fungus is selected from yeast (e.g., Debaryomyces hansenii ( Debaryomyces hansenii ), Aspergillus spp., Epidermophyton spp., or any combination thereof; Preferably, the disease is a disease caused by infection with Escherichia coli, Staphylococcus aureus and / or Debaryomyces hansenii; Preferably, the disease is selected from respiratory diseases (e.g., pneumonia, bronchitis), digestive system diseases (e.g., enteritis, gastritis, gastric ulcer, Crohn's disease), internal organ infections (e.g., meningitis, pericarditis), skin and soft tissue infections (e.g., folliculitis, furuncle, carbuncle), or any combination thereof.

9. Use of the coagulant Weizmannella according to any one of claims 1 to 3 or the composition according to claim 4 in preparing food; Preferably, the Weizmannella coagulans described in any one of claims 1 to 3 or the composition described in claim 4 is used as a starter for the food; Preferably, the food is solid food (eg, cheese, bread) or liquid food (eg, yogurt, flavored fermented milk, lactic acid bacteria beverage).

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