Lactobacillus plantarum and application thereof

Abstract

The invention discloses lactobacillus plantarum and application thereof, and aims at solving the technical problems of how to inhibit the growth of malassezia furfur, splitting the malassezia furfur, and inhibiting the growth of pseudomonas aeruginosa and staphylococcus aureus and the formation of a biological membrane. In particular discloses lactobacillus plantarum, the lactobacillus plantarum is lactobacillus plantarum DZ041, and the preservation number of the lactobacillus plantarum is CGMCC (China General Microbiological Culture Collection Center) No.20835 in the General Microbiological Culture Collection Center of the China Committee for Culture Collection of Microorganisms). Bacterial suspension or cell-free fermentation liquor prepared from the malassezia furfur can inhibit the growth of malassezia furfur and even can crack malassezia furfur, and the malassezia furfur can be used for industrial production.

Description

Technical Field

[0001] The invention specifically relates to a plant lactobacillus strain and application thereof. Background Art

[0002] As the soil for hair growth, the micro-ecological environment of hair is crucial. There are different types and numbers of microorganisms in various parts of human skin. Under normal conditions, these flora work together to maintain the health of the skin. When the flora is stimulated and unbalanced, skin diseases will occur. As a unique ecology, the human scalp has dense hair, many sweat glands, sebaceous glands and high relative humidity, which provide a suitable environment for the colonization and growth of microorganisms. The scalp microbiome is usually composed of bacteria and fungi, generally including Staphylococcus, Propionibacterium and Malassezia. The bacterial flora of the scalp is mainly Staphylococcus epidermidis, and the fungal flora is mainly Malassezia restricta and Malassezia furfur. Under normal living conditions, the scalp has a good metabolism and self-purification defense mechanism to inhibit the growth of scalp bacteria. A normal and healthy scalp is maintained by three major balances: oil, flora, and metabolism. The scalp is grayish white, with a clean surface, no dandruff, oily hair, redness, too thick stratum corneum, folliculitis, etc. When the balance is disrupted due to some external reasons (pollution, sun exposure, perming and dyeing) or internal reasons (diet, stress, endocrine disorders, bad living habits), various scalp problems will occur, such as dandruff, oily hair, folliculitis, etc. Therefore, care and treatment can be used to regulate the three major balances of the scalp and solve scalp health problems.

[0003] Malassezia is a resident flora in human skin and mucous membranes and is a lipophilic fungus. Different subspecies have different microscopic morphology, metabolic characteristics and genomic information. At present, 14 subspecies have been isolated and identified, including Malassezia spherical, Malassezia restricta, Malassezia furfur, Malassezia pachydermatis, etc. Malassezia furfur is a lipophilic yeast existing on the surface of the skin, accounting for 50-80% of the human skin fungal microbiome. In addition, they can form biofilms. Under normal symbiotic conditions, Malassezia mainly exists in the form of yeast. However, when the fungus is transformed into a pathogenic filamentous form, it can cause skin damage. Various Malassezia species exist as opportunistic pathogens and are also involved in the occurrence of various skin diseases, such as pityriasis versicolor (PV), Malassezia folliculitis (MF), dandruff / seborrheic dermatitis (D / SD), atopic dermatitis (AD) and psoriasis. Malassezia can participate in the occurrence of skin diseases through various pathways. The virulence factors of Malassezia include lipase, phospholipase, bioactive indole and other irritating Malassezia metabolites, and these extracellular macromolecules are involved in the development of the disease. In PV, indoles produced by Malassezia can mediate UV damage and melanin production by activating the aryl hydrocarbon receptor (AhR). Malassezia lipase promotes the occurrence of dandruff by locally producing irritating oleic acid, which in turn aggravates changes in skin barrier permeability. Studies have found that AD patients react more frequently to Malassezia extracts and recombinant Malassezia antigens. In psoriasis, Malassezia invades epidermal cells and makes keratinocytes more sensitive to stress responses, resulting in increased expression of antimicrobial peptides.

[0004] Lactobacillus plantarum is a type of lactic acid bacteria, with an optimal growth temperature of 30-35℃, anaerobic or facultative anaerobic, straight or curved rod-shaped, single, sometimes in pairs or chains, with an optimal pH of about 6.5, belonging to homofermentative lactic acid bacteria. The difference between this bacterium and other lactic acid bacteria is that the number of live bacteria is relatively high, and it can produce a large amount of acid, so that the pH value in the water is stable and does not rise. Lactobacillus plantarum has many health care effects, as follows: ① It has a certain immune regulation effect; ② It has an inhibitory effect on pathogenic bacteria; ③ It reduces serum cholesterol levels and prevents cardiovascular diseases; ④ It maintains the balance of intestinal flora; ⑤ It promotes the absorption of nutrients; ⑥ It relieves lactose intolerance; ⑦ It inhibits the formation of tumor cells, etc. Many strains of Lactobacillus plantarum have strong antagonistic activity against corruption, pathogenic bacteria and fungi. Recent studies have shown that Lactobacillus plantarum can produce different antimicrobial agents, such as organic acids, hydrogen peroxide, diacetyl and antimicrobial peptides, including bacteriocins. Its antimicrobial effects against pathogenic and spoilage microorganisms are generally attributed to the production of organic acids, including lactic acid and phenyllactic acid. Summary of the invention

[0005] The technical problem solved by the invention is how to provide an antibacterial plant lactobacillus.

[0006] In order to solve the above technical problems, the present invention provides Lactobacillus plantarum.

[0007] The plant lactobacillus is plant lactobacillus (Lactobacillus plantarum) DZ041, and its deposit number in the General Microbiological Center of China National Microbiological Culture Collection is CGMCC No.20835.

[0008] In order to solve the above technical problems, the present invention also provides a bacterial agent.

[0009] The bacterial agent contains the above-mentioned Lactobacillus plantarum.

[0010] The microbial agent has at least one of the following functions:

[0011] 1) inhibiting and / or slowing down the growth of Malassezia furfur; 2) preparing fermentation broth.

[0012] The active ingredient of the above-mentioned bacterial agent may be Lactobacillus plantarum DZ041, and the active ingredient of the above-mentioned bacterial agent may also contain other biological components or non-biological components. The other active ingredients of the above-mentioned bacterial agent may be determined by those skilled in the art according to the effects of the bacterial agent.

[0013] In order to solve the above technical problems, the present invention also provides a fermentation liquid.

[0014] The plant lactobacillus fermented liquid is obtained by fermenting the plant lactobacillus.

[0015] In the above, the molecular diameter of the substance in the fermentation broth is less than 0.22 μm.

[0016] In the above text, fermentation broth refers to liquid culture medium inoculated with microbial strains. After fermentation and cultivation, the microorganisms use the nutrients in the culture medium to synthesize bacteria and secrete products.

[0017] In the above, the fermentation step of the cell-free fermentation broth includes but is not limited to the following steps: culturing in MRS liquid culture medium at 28° C. or 37° C. for 24 h or 48 h, respectively.

[0018] In order to solve the above technical problems, the present invention also provides a composition.

[0019] The composition comprises the aforementioned Lactobacillus plantarum and / or the aforementioned bacterial agent and / or the aforementioned fermentation liquid.

[0020] In the above, the composition may further include a pharmaceutically acceptable excipient or a pharmaceutically acceptable carrier.

[0021] The above-mentioned plant lactobacillus is used in any of the following applications:

[0022] A1) Use in inhibiting or slowing the growth of fungi or bacteria and / or use in preparing products for inhibiting and / or slowing the growth of fungi or bacteria;

[0023] A2) Use in the lysis of fungi and / or in the preparation of lysis fungal products;

[0024] A3) Use in destroying the integrity of fungal cell membranes and / or use in preparing products that destroy the integrity of fungal cell membranes;

[0025] A4) Preparation for use in treating or assisting in treating diseases caused by fungi;

[0026] A5) Preparation for use in treating or assisting in treating diseases caused by fungi.

[0027] Application of the above-mentioned Lactobacillus plantarum in inhibiting bacterial biofilm formation or preparing products for inhibiting bacterial biofilm formation.

[0028] The use of the above-mentioned bacterial agent in any of the following:

[0029] A1) Use in inhibiting or slowing the growth of fungi and / or use in preparing products for inhibiting and / or slowing the growth of fungi;

[0030] A2) Use in the lysis of fungi and / or in the preparation of lysis fungal products;

[0031] A3) Use in destroying the integrity of fungal cell membranes and / or use in preparing products that destroy the integrity of fungal cell membranes;

[0032] A4) Preparation for use in treating or assisting in treating diseases caused by fungi;

[0033] A5) Preparation for use in treating or assisting in treating diseases caused by fungi;

[0034] A6) Use in inhibiting bacterial biofilm formation or in preparing products for inhibiting bacterial biofilm formation.

[0035] In the above, the fungus may be Malassezia furfur.

[0036] The use of the above-mentioned bacterial agent in inhibiting bacterial biofilm formation or preparing a product for inhibiting bacterial biofilm formation.

[0037] The fermentation broth is used in inhibiting or slowing the formation of fungal biofilms, in preparing products for inhibiting or slowing the formation of fungal biofilms, and / or in preparing products for treating or assisting in treating diseases caused by fungi.

[0038] In the above, the fungus may be Malassezia furfur, and the bacteria may be Pseudomonas aeruginosa and / or Staphylococcus aureus.

[0039] As in any of the above uses, the fungus is Malassezia furfur, Pseudomonas aeruginosa and / or Staphylococcus aureus.

[0040] In the above, the biofilm refers to a membrane-like multicellular complex formed by fungi attached to the host cavity or the surface of biological materials, secreting polysaccharide matrix, fibrin, lipoprotein, etc., and surrounding themselves therein. In the embodiment of the present application, it can be a biofilm of Staphylococcus aureus or Pseudomonas aeruginosa.

[0041] Beneficial Effects

[0042] The invention discloses a plant lactobacillus and its application, and solves the technical problem of how to inhibit the growth of Malassezia furfur, decompose Malassezia furfur or inhibit the formation of Malassezia furfur biomass. Specifically disclosed is a plant lactobacillus, which is Lactobacillus plantarum DZ041, and its deposit number in the General Microbiological Center of China Microbiological Culture Collection Administration Committee is CGMCC No.20835. The bacteria are prepared into a bacterial suspension or a cell-free fermentation liquid, which can inhibit the growth of Malassezia furfur and even decompose Malassezia furfur, and can be used for industrial production.

[0043] Collection Instructions

[0044] Bacteria name: Lactobacillus plantarum

[0045] Latin name: Lactobacillμsplantarμm

[0046] Strain ID: DZ041

[0047] Depository: China National Microbiological Culture Collection Administration General Microbiology Center

[0048] Abbreviation of depository institution: CGMCC

[0049] Address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing

[0050] Date of deposit: September 27, 2020

[0051] Registration number of the Collection Center: CGMCC No.20835 BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Figure 1 This is the detection diagram of DZ041 against Malassezia furfur; A is the positive control of Malassezia furfur; B is the inhibition zone of DZ041 against Malassezia furfur.

[0053] Figure 2This is the detection diagram of DZ041 CFS against Malassezia furfur; A. The inhibitory effect of different concentrations of CFS on M. furfur: Line 1-12: 1 (MRS + LN); 2 (MRS + M. furfur); 3 (90% CFS + M. furfur); 4 (80% CFS + M. furfur); 5 (70% CFS + M. furfur); 6 (60% CFS + M. furfur); 7 (50% CFS + M. furfur); 8 (40% CFS + M. furfur); 9 (30% CFS + M. furfur); 10 (20% CFS + M. furfur); 11 (10% CFS + M. furfur); 12 (100% CFS + M. furfur); B. The inhibition rate of different concentrations of CFS on M. furfur.

[0054] Figure 3 Scanning electron micrographs (SEM) of the effects of Lactobacillus plantarum DZ041 CFS on Malassezia furfur (M. furfur); A: MRS + M. furfur; B: CFS (KB) + M. furfur; C: 100% CFS (DZ041) + M. furfur; D: 50% CFS (DZ041) + M. furfur; E: 40% CFS (DZ041) + M. furfur.

[0055] Figure 4 These are the confocal fluorescence microscopy results of the effect of the supernatant of Lactobacillus plantarum DZ041 on Malassezia furfur (M. furfur).

[0056] Figure 5 This is a graph showing the inhibitory effect of Lactobacillus plantarum DZ041 on M. furfur at different culture temperatures and times.

[0057] Figure 6 This is a graph showing the effect of Lactobacillus plantarum DZ041 cell-free fermentation broth (CFS) on the biofilm formation and mature biofilm removal of S. aureus and P. aeruginosa; Contrast is S. aureus / P. aeruginosa+MRS; Exp1 is S. aureus / P. aeruginosa+CFS(DZ041); Exp2 is S. aureus / P. aeruginosa mature biofilm+CFS(DZ041). Figure 7 This is a graph showing the inhibitory effect of different concentrations of CFS on the growth of S. aureus and P. aeruginosa.

[0058] Figure 8This is a graph showing the inhibitory effect of Lactobacillus plantarum DZ041 CFS on the growth of Pseudomonas aeruginosa and Staphylococcus aureus; S. aureus is Staphylococcus aureus, P. aeruginosa is Pseudomonas aeruginosa; the ordinate is the inhibition rate; the abscissas 5%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, and 50% are the final volume fractions of CFS of Lactobacillus plantarum DZ041, respectively. DETAILED DESCRIPTION

[0059] The present invention is further described in detail below in conjunction with specific embodiments, and the examples provided are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way.

[0060] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0061] The following examples used SPSS11.5 statistical software to process the data, and the experimental results were expressed as mean ± standard deviation. One-way ANOVA test was used, and P < 0.05 (*) indicated a significant difference, P < 0.01 (**) indicated a very significant difference, and P < 0.001 (***) indicated an extremely significant difference.

[0062] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this article.

[0063] In order to make the present invention more easily understood, certain technical and scientific terms are specifically defined below. Unless otherwise clearly defined elsewhere in this document, all other technical and scientific terms used herein have the meanings commonly understood by those skilled in the art to which the present invention belongs.

[0064] The terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of the features. In the description of the present invention, the meaning of "plurality" and "multiple" is at least two or two, such as two, two, three, three, etc., unless otherwise clearly and specifically defined.

[0065] The terms “include”, “have” or “comprises” are open expressions, that is, including the contents specified in the present invention but not excluding other contents.

[0066] The term "microbial agent" refers to a prepared live bacterial preparation, which may include a carrier (solid carrier or liquid carrier) and an auxiliary material. The microbial agent may be in a variety of dosage forms, including but not limited to liquid, emulsion, suspension, powder, granule, wettable powder or water-dispersible granule. The solid carrier may be a mineral material, a plant material or a polymer compound; the mineral material may be at least one of clay, talc, kaolin, montmorillonite, white carbon, zeolite, silica and diatomaceous earth; the plant material may be at least one of corn flour, soybean flour and starch; the polymer compound may be polyvinyl alcohol and / or polyglycol. The liquid carrier may be vegetable oil, mineral oil or water; the organic solvent may be decane and / or dodecane. In the microbial agent, the active ingredient may be in the form of cultured living cells, fermentation broth of living cells, filtrate of cell culture or a mixture of cells and filtrate. The auxiliary materials include water, carbon source and / or ammonia source. Carbon source is a type of nutrient for microbial growth, which is a carbon-containing compound, including fast-acting and slow-acting carbon sources such as sugars, oils, organic acids and organic acid esters and small molecule alcohols. Hydrogen source refers to a substance that provides nitrogen elements required for microbial nutrition, including peanut cake powder, soybean cake powder, yeast powder, peptone, ammonia water, ammonium salts and nitrates and other fast-acting and slow-acting hydrogen sources. The dosage form of the bacterial agent can be a variety of dosage forms, such as liquid, suspension, powder, granule, wettable powder or water-dispersible granule. In the present application, the active ingredient of the bacterial agent can be Lactobacillus plantarum DZ041, and the active ingredient of the bacterial agent can also contain other biological components or non-biological components. The other active ingredients of the above-mentioned bacterial agent can be determined by those skilled in the art according to the effect of the bacterial agent.

[0067] The term "fermentation broth" refers to a solution obtained after culturing Lactobacillus plantarum DZ041 for a period of time, which solution mainly contains Lactobacillus plantarum DZ041 and its metabolites; or a supernatant after further treatment by means of centrifugation, filtration, etc., which supernatant mainly contains metabolites of Lactobacillus plantarum DZ041.

[0068] The term "treatment" refers to the use of drugs to obtain the desired pharmacological and / or physiological effects. The effect may be preventive in terms of completely or partially preventing a disease or its symptoms, and / or may be therapeutic in terms of partially or completely curing a disease and / or the adverse effects caused by the disease. "Treatment" as used herein covers diseases in mammals, especially humans, including: (a) preventing the occurrence of a disease or condition in individuals who are susceptible to the disease but have not yet been diagnosed with the disease; (b) inhibiting the disease, such as blocking the progression of the disease; or (c) alleviating the disease, such as alleviating symptoms associated with the disease. "Treatment" as used herein covers any medication that administers a strain, fermentation broth or composition to an individual to treat, cure, alleviate, improve, reduce or inhibit the individual's disease, including but not limited to administering a strain, fermentation broth or composition described herein to an individual in need.

[0069] The term "carrier" includes any solvent, drug stabilizer, or combination thereof, which are known to those skilled in the art. Except in the case where any conventional carrier is incompatible with the active ingredient, its use in treatment or pharmaceutical composition is covered.

[0070] In the present application, "Lactobacillus plantarum DZ041", "Lactobacillus plantarum DZ041", "DZ041", "Lactobacillus plantarum DZ041", "CGMCC No. 20835", "DZ041 strain" or "strain DZ041" are synonymous.

[0071] The term "pharmaceutically acceptable carrier" includes any solvent, drug stabilizer, or combination thereof, which are known to those skilled in the art. Except in the case where any conventional carrier is incompatible with the active ingredient, its use in treatment or pharmaceutical composition is covered.

[0072] The term "pharmaceutically acceptable excipient" is a pharmaceutically or food-acceptable carrier, solvent, suspending agent or excipient for delivering the active ingredient in the medicine of the present invention to an animal or human. Exemplary excipients can be liquid or solid, including but not limited to: pH regulators, surfactants, carbohydrates, adjuvants, antioxidants, chelating agents, ionic strength enhancers, preservatives, carriers, glidants, sweeteners, dyes / colorants, flavor enhancers, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents, emulsifiers, sprays, compressed air or other suitable gases, or other suitable inactive ingredients used in combination with pharmacodynamic compounds. Examples of excipients include various lactose, mannitol, oils such as corn oil, buffers such as PBS, saline, polyethylene glycol, glycerol, polypropylene glycol, dimethyl sulfoxide, amides such as dimethylacetamide, proteins such as albumin, monosaccharides and oligosaccharides such as glucose, lactose, cyclodextrin and starch.

[0073] The term "therapeutically effective amount" or "therapeutically effective dose" refers to an amount of a substance, compound, material, or composition comprising a compound that is at least sufficient to produce a therapeutic effect after administration to a subject. Therefore, it is the amount necessary to prevent, cure, improve, block, or partially block the symptoms of a disease or disorder.

[0074] In the present application, Malassezia furfur is Malassezia furfur, which can cause tinea versicolor, pityrosporum pilaris, dandruff, seborrheic dermatitis, atopic dermatitis. Pseudomonas aeruginosa is Pseudomonas aeruginosa. Staphylococcus aureus is Staphylococcus aureus.

[0075] Acquisition of Malassezia furfur

[0076] The strain number of Malassezia furfur is M. furfur ATCC 44344.

[0077] Obtaining Lactobacillus plantarum DZ041

[0078] The strain of the present invention is isolated from the scalp of a healthy person. A sterile cotton swab for collecting a scalp sample is added to sterile physiological saline and shaken and mixed, and 100 μL is spread on an MRS agar plate. After constant temperature anaerobic static culture at 37° C. for 24 hours, strains with different colony morphologies are selected, and streaking purification culture is performed on the plate again to obtain isolated strains.

[0079] The genomic DNA of DZ041 was extracted with the Omega kit, and the 16S rRNA sequence was amplified with universal primers 27f and 1492r. After gel excision and purification, the pMD-18T vector was connected. The positive colonies verified by colony PCR were inoculated in liquid LB medium (5g yeast extract, 10g tryptone, 5g sodium chloride, 18g agar, pH7.4) with Amp (final concentration of 50μg / mL). The bacterial liquid after overnight culture was sent to Nosai Biotechnology for sequencing. The sequencing results were spliced ​​by DNAMAN software and submitted to http: / / www.ezbiocloud.net / eztaxon for sequence analysis. The sequence alignment results showed that DZ041 belonged to the genus Lactobacillus plantarum, and it was named Lactobacillus plantarum DZ041 or Lactobacillus plantarum DZ041. The above-mentioned target strain was deposited in the General Microbiology Center of China Microbiological Culture Collection through patent procedures on September 27, 2020. The deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No. 20835.

[0080] The 16S rRNA sequence is shown in Sequence 1, and is as follows:

[0081]

[0082] Example 1 Detection of the ability of Lactobacillus plantarum DZ041 to resist Malassezia furfur

[0083] MRS liquid culture medium: The solutes and their concentrations of MRS liquid culture medium are peptone 10g / L, gravy 8g / L, yeast powder 4g / L, glucose, 20g / L, sodium acetate 5g / L, Tween 801g / L, disodium hydrogen phosphate 2g / L, triammonium citrate 2g / L, magnesium sulfate 0.2g / L, manganese sulfate 0.05g / L, the solvent is water, pH 6.2.

[0084] Leeming-Notman (LN) liquid medium: peptone 10.0g / L, glucose 5.0g / L, yeast extract powder 1.0g / L, ox bile salt 4.0g / L, glycerol 1.0g / L, monostearate glyceryl 0.5g / L, Tween 60 0.5g / L, whole milk powder 10.0g / L, chloramphenicol 0.05g / L, pH 5.6±0.2 (25℃), weigh 32.0g of this product in 1L distilled water or deionized water, heat and boil to dissolve, dispense into triangular bottles (lay a layer of glass beads on the bottom of the bottle), add 1.6ml olive oil to every 80ml, sterilize at 121℃ for 15 minutes, cool to about 50℃, add 1 bottle of S0614 to every 80ml aseptically, shake well, and set aside. Purchased from Shandong Topological Bioengineering Co., Ltd.

[0085] MRS solid plate: Add 2g / L agar powder to MRS liquid medium.

[0086] Leeming-Notman agar medium: Add 2 g / L agar powder to LN liquid medium.

[0087] 1. A single clone of Lactobacillus plantarum DZ041 was inoculated into a test tube containing 5 mL of MRS liquid culture medium and cultured at 37° C. for 24 h. The culture medium was the fermentation medium of Lactobacillus plantarum DZ041.

[0088] 2. Inoculate a single clone of Malassezia furfur (M. furfur) into a test tube containing 5 mL of Leeming-Notman (LN) liquid culture medium, and culture at 30° C., 200 rpm / min for 2 days to obtain a Malassezia furfur suspension.

[0089] 3. Add 5 μL of Lactobacillus plantarum DZ041 fermentation liquid to the MRS solid plate at 37°C, and culture at constant temperature for 24 hours. Inject 1 mL of Malassezia furfur suspension into 80 mL of unsolidified Leeming-Notman agar medium, and pour 15 mL to the upper layer of the MRS solid plate. Culture at 30°C aerobically for 5 days, and observe the size of the transparent circle. The plate without adding Lactobacillus plantarum DZ041 fermentation liquid was used as a control. According to the transparent circle, it can be seen that DZ041 significantly inhibits the growth of Malassezia furfur. Results are shown in Figure 1 As shown, the results showed that DZ041 could inhibit the growth of Malassezia furfur (M. furfur).

[0090] Example 2 Detection of the ability of Lactobacillus plantarum DZ041 CFS to fight Malassezia furfur

[0091] The frozen strain of Lactobacillus plantarum DZ041 was inoculated into 5 mL of MRS liquid culture medium, and cultured at 37° C. for 24 h to obtain bacterial solution 1. The bacterial solution was streaked on an MRS plate (MRS solid plate), and cultured at 37° C. for 24 h. A single clone was picked and inoculated into 10 mL of MRS liquid culture medium, and cultured at 37° C. for 48 h to obtain bacterial solution 2. Bacterial solution 2 was centrifuged at 8000 rpm / min, 4° C. for 15 min to obtain a supernatant. The supernatant was filtered through a 0.22 μm PES (polyethersulfone resin) membrane filter to obtain a cell-free fermentation solution (CFS) of Lactobacillus plantarum DZ04.

[0092] The cell-free fermentation broth (CFS) of Lactobacillus plantarum DZ041 was diluted with sterile MRS liquid medium to a volume fraction (i.e., concentration) of 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, and 10% of the cell-free fermentation broth (CFS) of Lactobacillus plantarum DZ041. The above dilutions were mixed with the Malassezia furfur suspension at a volume ratio of v:v=1:1 and then inoculated into a sterile 96-well polypropylene plate. The final concentration of the Malassezia furfur suspension was 1×10 7 cfu / mL, the volume proportion of CFS in the mixed system (i.e., final concentration) was 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, and 5%. The positive control group (OD1) was sterile MRS liquid medium + an equal volume of Malassezia furfur suspension (the final concentration of Malassezia furfur suspension was 1×10 7 cfu / mL), the negative control group (OD0) was sterile MRS liquid medium + Leeming-Notman liquid medium, the 96-well plate was cultured at 30°C for 48 h, the OD595 was measured by a microplate reader, and the inhibition rate was calculated.

[0093] Inhibition rate = [1-(ODt-OD0) / (OD1-OD0)] × 100%

[0094] ODt: CFS+M.furfur; OD0: MRS+LN; OD1: MRS+M.furfur

[0095] Results Figure 2 , the diluted cell-free fermentation broth (CFS) of Lactobacillus plantarum DZ041 can still inhibit the growth of Malassezia furfur, and the minimum inhibitory concentration is about 25%. The inhibitory effect of CFS is concentration-dependent. After 48 hours of co-culture, 50% CFS (final concentration of 25%) is the minimum inhibitory concentration of Malassezia furfur, and there is almost no inhibitory effect below this concentration.

[0096] Example 3 Effect of Lactobacillus plantarum DZ041 CFS on the surface structure of Malassezia furfur

[0097] The present invention isolates a strain from Korean kimchi, and the 16S rRNA sequence is detected to be shown in Sequence 2. It is found to be Leuconostoc mesenteroides, and is named strain KB.

[0098] Sequence 2 is as follows:

[0099]

[0100] LN liquid culture medium: peptone 10.0g / L, glucose 5.0g / L, yeast extract powder 1.0g / L, ox bile salt 4.0g / L, glycerol 1.0g / L, monostearate glyceryl 0.5g / L, Tween 60 0.5g / L, whole milk powder 10.0g / L, chloramphenicol 0.05g / L, pH 5.6±0.2 (25℃) Weigh 32.0g of this product in 1L distilled water or deionized water, heat and boil to dissolve, dispense into triangular bottles (lay a layer of glass beads on the bottom of the bottle), add 1.6ml olive oil to every 80ml, sterilize at 121℃ for 15 minutes, cool to about 50℃, add 1 bottle of S0614 to every 80ml aseptically, shake well, and set aside. Purchased from Shandong Topological Bioengineering Co., Ltd.

[0101] Diluted M. furfur: Dilute M. furfur with sterile LN.

[0102] The frozen strain of Malassezia furfur was inoculated into a test tube containing 5 mL of LN liquid culture medium at 5%, and cultured in a shaker at 30 ° C, 200 rpm / min for 48 h. The OD600 was measured by UV spectrophotometer, and the LN liquid culture medium was diluted to a concentration of 2 × 10 7 cfu / mL, Lactobacillus plantarum DZ041 and strain KB were inoculated in test tubes containing 8 mL of MRS liquid culture medium, cultured at 37°C for 24 h, centrifuged at 8000 rpm / min for 10 min, and the supernatant was filtered through a 0.22 μm PES filter to obtain CFS of Lactobacillus plantarum DZ041 and CFS of strain KB. CFS of Lactobacillus plantarum DZ041 (100%, 50%, 40%) and CFS of strain KB (100%) were mixed with Malassezia furfur (M. furfur) in a volume ratio of 1:1 in a test tube, so that the final concentration of Malassezia furfur (M. furfur) was 1×10 7 cfu / mL, shaker at 30°C, 200rpm / min, co-culture for 48h, control group is MRS (100%) and Malassezia furfur (M. furfur). After culture, the bacterial solution is centrifuged at 6000rpm / min for 5min, the supernatant is discarded to obtain the bacteria, and the same volume of sterile PBS (pH=7.5) is resuspended and then centrifuged again, and repeated three times. After centrifugation, the bacteria are fixed with the same volume of 2.5% glutaraldehyde at 4°C, dark for 12h, 6000rpm / min, centrifuged for 5min, the supernatant is discarded, the bacterial precipitation is successively mixed with 30%, 50%, 70%, 90% ethanol by blowing and mixing, soaked for 10min, and then centrifuged for 5min, and finally the same volume of anhydrous ethanol is used to dissolve the precipitation, 3-5μL is coated on a 10×10mm silicon wafer, and naturally air-dried in a clean bench, and the morphological characteristics of the bacteria are observed by scanning electron microscopy (SEM).

[0103] The results are as follows Figure 3 (A is the control group, B is CFS (KB) + M. furfur, C is 100% CFS (DZ041) + M. furfur; D is 50% CFS (DZ041) + M. furfur, and E is 40% CFS (DZ041) + M. furfur), inoculated in MRS liquid culture medium at 37°C, cultured for 24 hours, and the pH of the fermentation liquid was measured to be <5. KB was used as a positive control, and the culture conditions were the same as those of DZ041. The bacterial morphology of the CFS (KB) and 40% CFS (DZ041) treatment groups was the same as that of the control group, indicating that there was no inhibitory effect on the bacteria, while the bacteria in the 100% CFS and 50% CFS treatment groups ruptured and the contents leaked, which was consistent with Figure 2 The minimum inhibitory effect is shown in Table 1. The total incubation time of the treatment group was 48 hours, at which time most of the bacteria had died.

[0104] Example 4 Effect of CFS of Lactobacillus plantarum DZ041 on the cell membrane integrity of Malassezia furfur

[0105] The frozen strain of Malassezia furfur was inoculated into a test tube containing 5 mL of LN liquid culture medium at 5%, and cultured in a shaker at 30 ° C, 200 rpm / min for 48 h. The OD600 was measured by UV spectrophotometer, and the LN liquid culture medium was diluted to a concentration of 2 × 10 7 cfu / mL, plant lactobacillus DZ041 and strain KB were inoculated in test tubes containing 8 mL MRS liquid culture medium, cultured at 37° C. for 24 h, centrifuged at 8000 rpm / min for 10 min, and the supernatant was filtered through a 0.22 μm filter membrane to obtain CFS of plant lactobacillus DZ041 and CFS of strain KB. The CFS of plant lactobacillus DZ041 was diluted with MRS to obtain undiluted 100% CFS (DZ041), 50% CFS (DZ041) and 40% CFS (DZ041), while the CFS of strain KB was not diluted to obtain undiluted 100% CFS (KB). 100% CFS (KB), 100% CFS (DZ041), 50% CFS (DZ041), and 40% CFS (DZ041) were mixed with Malassezia furfur (M. furfur) at a volume ratio of 1:1 (3 mL + 3 mL) in a test tube. The final concentrations of CFS were 50% CFS (KB), 50% CFS (DZ041), 25% CFS (DZ041), and 20% CFS (DZ041), and the final concentration of Malassezia furfur (M. furfur) was 1×10 7cfu / mL, the test tubes were placed in a shaker at 30°C, 200 rpm / min, and cultured for 48 h. The control group was undiluted MRS mixed with Malassezia furfur in a volume ratio of 1:1, and the final concentration of MRS was 50%. The cultured bacterial solution was centrifuged at 6000 rpm / min for 5 min, the supernatant was discarded, and the cells were resuspended with the same volume of sterile PBS (pH = 7.5) and centrifuged again, repeated three times, the cells were treated with 200 μL of propidium iodide PI (red fluorescent dye) with a final concentration of 2 mg / mL and 20 μM SYTO9 (green fluorescent dye) at 30°C, incubated for 2 h in the dark, centrifuged at 6000 rpm / min for 5 min, the dye was removed, and the cells were resuspended with 1 mL of sterile PBS, centrifuged at 6000 rpm / min for 5 min, the supernatant was discarded, and the washing was repeated 8-10 times until no red fluorescent dye remained in the background, 7 μL of the sample was dropped on a slide, covered with a coverslip, pressed gently, and the staining results were observed under a confocal laser scanning microscope (CLSM).

[0106] The staining results are as follows Figure 4 ( Figure 4 In the figure, *% represents the final concentration compared with the original concentration, CFS is cell-free fermentation broth, KB is strain KB, and DZ041 is Lactobacillus plantarum DZ041; from left to right, the first case is natural light, the second column is the superposition of green fluorescence and red fluorescence, the third column is green fluorescence, and the fourth column is red fluorescence). As shown in the figure, CFS (DZ041) has a good destructive effect on M. furfur yeast cells, and KB has no effect. SYTO9 can enter cells with intact and damaged cell membranes, while PI can only penetrate into cells with damaged cell membranes, causing them to produce green and red fluorescence respectively under the corresponding wavelength excitation light. The fluorescence intensity varies with the degree of membrane damage. The more severe the damage, the stronger the fluorescence intensity. The bacteria in the positive control group were not damaged, the cell membrane was intact, and PI could not enter the cells, so there was no red fluorescence. The 50% CFS (DZ041) group had a stronger ability to damage the cell membrane of the yeast phase of M. furfur, and the fluorescence intensity was the strongest. The 25% CFS (DZ041) group still had cell membrane damage, but the degree of damage was weakened, so the fluorescence intensity decreased accordingly. When the concentration of CFS (DZ041) was reduced to 20%, it had almost no destructive effect on the fungal cell membrane, and there was no red fluorescence. 50% CFS (KB) had no destructive effect on the fungal cell membrane.

[0107] Example 5 Inhibitory effect of Lactobacillus plantarum DZ041 CFS on Malassezia furfur under different conditions

[0108] Lactobacillus plantarum DZ041 was inoculated into a test tube containing 8 mL of MRS liquid culture medium at a ratio of 1%, and cultured at 28° C. and 37° C. for 24 h and 48 h, respectively, at 8000 rpm / min, 4° C., centrifuged for 10 min, and filtered through a 0.22 μm filter membrane to obtain CFS (cell-free fermentation broth of Lactobacillus plantarum DZ041), which was divided into 4 groups, namely, a 37° C. / 24 h group, a 37° C. / 48 h group, a 28° C. / 24 h group, and a 28° C. / 48 h group (the 37° C. / 24 h group was added with the above-mentioned Lactobacillus plantarum DZ041 cultured at 37° C. for 24 h to obtain the Lactobacillus plantarum DZ041 obtained by constant temperature static culture at 37° C. The 37°C / 48h group was added with the Lactobacillus plantarum DZ041 cell-free fermentation broth obtained by constant temperature static culture of the Lactobacillus plantarum DZ041 at 37°C for 48h, the 28°C / 24h group was added with the Lactobacillus plantarum DZ041 cell-free fermentation broth obtained by constant temperature static culture of the Lactobacillus plantarum DZ041 at 28°C for 24h, and the 28°C / 48h group was added with the Lactobacillus plantarum DZ041 cell-free fermentation broth obtained by constant temperature static culture of the Lactobacillus plantarum DZ041 at 28°C for 48h). 50 μL of CFS of the above four groups of Lactobacillus plantarum DZ041 with different culture times and temperatures were mixed with 150 μL M.furfur and added to a 96-well plate. The final concentration of M.furfur was 1×10 7 Each group had 6 replicate wells, the negative control group (OD0) was 50μL MRS + 150μL LN, the positive control group (OD1) was 50μL MRS + 150μL M.furfur, after 48h of constant temperature static culture at 28℃, OD595 was measured by microplate reader, and the inhibition rate was calculated.

[0109] Inhibition rate = [1-(ODt-OD0) / (OD1-OD0)] × 100%;

[0110] ODt: CFS+M.furfur; OD0: MRS+LN; OD1: MRS+M.furfur

[0111] The results are as follows Figure 5 ( Figure 5 In the figure, the horizontal axis: 37℃ / 24h is the 37℃ / 24h group, 37℃ / 48h is the 37℃ / 48h group, 28℃ / 24h is the 28℃ / 24h group, and 28℃ / 48h is the 28℃ / 48h group; the vertical axis: inhibition rate) shows that the inhibition effect of culturing for 24h is poor, and the production of bacterial metabolites is incomplete. The inhibition after culturing for 48h is stronger and is basically not affected by the two culture temperatures shown in the experiment. The amount of metabolites inhibiting Malassezia furfur (M. furfur) produced at the two temperatures for 48h is basically the same.

[0112] Example 6 Inhibitory effect of Lactobacillus plantarum DZ041 CFS on biofilm

[0113] TSB liquid culture medium: tryptone 17 g / L, soy peptone 3 g / L, sodium chloride 5 g / L, dipotassium hydrogen phosphate 2.5 g / L, glucose 2.5 g / L, pH 7.3±0.2, 25°C.

[0114] Pseudomonas aeruginosa liquid: a mixture of Pseudomonas aeruginosa activated for 8-10 hours and TSB medium

[0115] Staphylococcus aureus liquid: a mixture of Staphylococcus aureus activated for 8-10 hours and TSB medium

[0116] Staphylococcus aureus: S.aureus ATCC 12600

[0117] Pseudomonas aeruginosa: P. aeruginosa ATCC PA01

[0118] The biofilm formation inhibition experiment was performed as follows:

[0119] Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa) were inoculated into TSB liquid culture medium, shaken at 37°C, 200 rpm / min, and activated for 8-10 h to obtain activated Staphylococcus aureus (S. aureus) bacterial solution A and activated Pseudomonas aeruginosa (P. aeruginosa) bacterial solution A. The two bacterial solutions A were diluted with fresh sterile TSB to obtain Staphylococcus aureus bacterial solution B with OD600 = 0.08 and Pseudomonas aeruginosa bacterial solution B with OD600 = 0.04. The 12-well polypropylene plate was divided into two groups, 1 mL of Staphylococcus aureus bacterial solution B was added to each well of one group, and 1 mL of Pseudomonas aeruginosa bacterial solution B was added to each well of the other group, and then 1 mL of CFS of Lactobacillus plantarum DZ041 prepared above was added respectively. The final OD600 of Staphylococcus aureus and Pseudomonas aeruginosa were 0.04 and 0.02, respectively, to obtain S. aureus. Exp1 group and P.aeruginosa Exp1 group, the control group was divided into two groups in a 12-well polypropylene plate, 1 mL of Staphylococcus aureus liquid B was added to each well of one group, and 1 mL of Pseudomonas aeruginosa liquid B was added to each well of the other group, and then 1 mL of the MRS liquid culture medium prepared above was added respectively, so that the final OD600 of Staphylococcus aureus and Pseudomonas aeruginosa were 0.04 and 0.02, respectively, to obtain the S.aureus Contrast group and the P.aeruginosa Contrast group, insert a 18 mm × 18 mm cover glass into each well, and culture at 28 ° C at a constant temperature and aerobic static culture for 46-48 h until a biofilm was formed on the cover glass and the inner wall of the well, aspirate all the culture solution in the well, slowly rinse the cover glass with sterile PBS (pH 7.5) to remove the floating bacteria, add 2 mL of methanol to each well for immersion and fixation for 20 min, aspirate all the methanol, rinse the cover glass and the well with PBS to remove the residual methanol, and add 2 mL of Stain with 0.01% crystal violet at room temperature in the dark for 20 min, aspirate the crystal violet solution, rinse the coverslip and the wells with PBS to remove residual crystal violet, air-dry at room temperature, remove the coverslip and observe the biofilm removal effect under an optical microscope at 40×.

[0120] The removal experiment of mature biofilm is as follows:

[0121] Staphylococcus aureus (S.aureus) and Pseudomonas aeruginosa (P.aeruginosa) were inoculated into TSB liquid culture medium, shaken at 37°C, 200 rpm / min, and activated for 8-10 hours to obtain activated Staphylococcus aureus (S.aureus) bacterial solution A and activated Pseudomonas aeruginosa (P.aeruginosa) bacterial solution A. The two bacterial solutions A were diluted with fresh sterile TSB to obtain Staphylococcus aureus bacterial solution B with OD600=0.04 and Pseudomonas aeruginosa bacterial solution B with OD600=0.02, and divided into two groups in a 12-well polypropylene plate, 2 mL of Staphylococcus aureus bacterial solution B was added to each well of one group, and 2 mL of Pseudomonas aeruginosa bacterial solution B was added to each well of the other group. The final OD600 of Staphylococcus aureus and Pseudomonas aeruginosa in bacterial solution B were 0.04 and 0.02, respectively, to obtain S.aureus Exp2 group and P.aeruginosa In the Exp2 group, a 18 mm × 18 mm coverslip was inserted into each well and cultured aerobically at 28°C for 46-48 h until a mature biofilm formed on the coverslip and the inner wall of the well. The culture medium in the well was aspirated and the planktonic bacteria on the inner wall of the well and the coverslip were rinsed with sterile PBS. 2 mL of CFS (final concentration 100%) was added to each well and incubated at 28°C for 24 h. All CFS in the well was aspirated and the coverslip was slowly rinsed with sterile PBS (pH 7.5) to remove the planktonic bacteria. 2 mL of methanol was added to each well for immersion and fixation for 20 min. All methanol was aspirated and the coverslip and the well were rinsed with PBS to remove the residual methanol. 2 mL of 0.01% crystal violet was added to each well for staining at room temperature and away from light for 20 min. The crystal violet solution was aspirated and the coverslip and the well were rinsed with PBS to remove the residual crystal violet. After air drying at room temperature, the coverslip was removed and the biofilm removal effect was observed under an optical microscope at 40×. ( Figure 6 )

[0122] The results are as follows Figure 6 ( Figure 6 (in the figure, S. aureus is Staphylococcus aureus, P. aeruginosa is Pseudomonas aeruginosa, Contrast is control, EXP1 is the biofilm formation inhibition experiment, and EXP2 is the biofilm removal experiment) As shown in the figure, the CFS of Lactobacillus plantarum DZ041 can inhibit the formation of S. aureus and P. aeruginosa biofilms, and has basically no removal effect on mature biofilms.

[0123] Example 7 Inhibition rate of biofilm formation by Lactobacillus plantarum DZ041 CFS

[0124] The biofilm formation inhibition rate was determined as follows:

[0125] Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa) were inoculated into TSB liquid culture medium, shaken at 37°C, 200 rpm / min, and activated for 8-10 hours to obtain activated Staphylococcus aureus (S. aureus) bacterial solution A and activated Pseudomonas aeruginosa (P. aeruginosa) bacterial solution A. The two bacterial solutions A were diluted with fresh sterile TSB to obtain Staphylococcus aureus bacterial solution B with OD600 = 0.08 and Pseudomonas aeruginosa bacterial solution B with OD600 = 0.04. The CFS of Lactobacillus plantarum DZ041 prepared above was diluted to Lactobacillus plantarum DZ041 with sterile MRS according to the volume fraction. The volume proportion (concentration) of CFS was 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, and 10%. CFS was mixed with S. aureus and P. aeruginosa at a volume ratio of 1:1, respectively, and inoculated in a 96-well plate, 200uL per well. The final concentrations of CFS were 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, and 5% respectively. The concentrations of S. aureus and P. aeruginosa in the wells were 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, and 5%. The final concentrations of P.aeruginosa and P.aeruginosa were OD600 = 0.04 and OD600 = 0.02, respectively. The cells were incubated in a constant temperature incubator at 28 °C for 48 h. The culture medium in the wells was aspirated, and 200 μL of sterile PBS was added to each well. The cells were washed 2-3 times to remove floating bacteria. The PBS in the wells was aspirated, and 200 μL of methanol was added to each well to fix at room temperature for 20 min. The methanol in the wells was aspirated, and an equal amount of sterile PBS was added to wash twice. After aspirating PBS, 200 μL of 0.01% crystal violet solution was added to each well. The cells were incubated at room temperature and away from light for 20 min. The crystal violet solution was aspirated, and 200 μL of PBS was added to each well to wash 2-3 times to remove the residual crystal violet solution in the wells. After natural air drying at room temperature, 200 μL of acetic acid solution with a volume fraction of 33% was added to each well to dissolve the biofilm remaining on the inner wall. The OD595 was measured by an enzyme marker. The inhibition rate of biofilm formation of P.aeruginosa and S.aureus was determined as described above.

[0126] Inhibition rate = [1-(ODt-OD0) / (OD1-OD0)] × 100%;

[0127] OD0: 100μL TSB + 100μL MRS; OD1: 100μL S.aureus / P.aeruginosa bacterial solution B + 100μMRS; ODt: 100μL S.aureus / P.aeruginosa bacterial solution B + 100μL CFS.

[0128] The results are as follows Figure 7 ( Figure 7In the figure, S.aureus is Staphylococcus aureus, P.aeruginosa is Pseudomonas aeruginosa; the ordinate is the inhibition rate; the abscissa is 5%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% respectively, which are the final volume fractions of CFS of Lactobacillus plantarum DZ041. The minimum inhibition concentrations of CFS of Lactobacillus plantarum DZ041 on biofilm formation of Staphylococcus aureus and Pseudomonas aeruginosa are 20% and 5%, respectively. Compared with Staphylococcus aureus, CFS (DZ041) has a more significant inhibitory effect on biofilm formation of Pseudomonas aeruginosa, and still has a strong inhibitory effect at lower concentrations.

[0129] Example 8 Inhibitory Effect of Lactobacillus plantarum DZ041 CFS on the Growth of Pseudomonas aeruginosa and Staphylococcus aureus

[0130] Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa) were inoculated into TSB liquid culture medium, shaken at 37°C, 200 rpm / min, and activated for 8-10 h to obtain activated Staphylococcus aureus (S. aureus) bacterial solution A and activated Pseudomonas aeruginosa (P. aeruginosa) bacterial solution A. The two bacterial solutions A were diluted with fresh sterile TSB to obtain Staphylococcus aureus bacterial solution B and Pseudomonas aeruginosa bacterial solution B. The concentration of bacterial solution B was 2×10 7 cfu / mL. The CFS of Lactobacillus plantarum DZ041 prepared above was diluted with sterile MRS according to volume fraction to a volume ratio (concentration) of 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, and 10% of Lactobacillus plantarum DZ041 CFS, respectively. CFS was mixed with S. aureus and P. aeruginosa bacterial solution B at a volume ratio of 1:1, respectively, and inoculated into 96-well plates, 200 μL per well, and the final concentrations of CFS were 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, and 5%, respectively. The final concentrations of S. aureus and P. aeruginosa in the wells were 1×10 7 cfu / mL, incubated in a 28°C constant temperature incubator for 24 h, measured OD595 with an ELISA reader, and calculated the inhibition rate.

[0131] Inhibition rate = [1-(ODt-OD0) / (OD1-OD0)] × 100%;

[0132] OD0: 100μLTSB+100μLMRS; OD1: 100μL S.aureus / P.aeruginosa bacterial solution B+100μL MRS; ODt: 100μL S.aureus / P.aeruginosa bacterial solution B+100μL CFS.

[0133] The results are as follows Figure 8 As shown, ( Figure 8 In the figure, S.aureus is Staphylococcus aureus, P.aeruginosa is Pseudomonas aeruginosa; the ordinate is the inhibition rate; the abscissa is 5%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, respectively, which are the final volume fractions of CFS of Lactobacillus plantarum DZ041). The minimum inhibitory concentrations of CFS of Lactobacillus plantarum DZ041 on the growth of bacteria in Staphylococcus aureus culture B and Pseudomonas aeruginosa culture B are 15% and 10%, respectively. Compared with Staphylococcus aureus, CFS (DZ041) has a more significant inhibitory effect on the growth of Pseudomonas aeruginosa, and still has a strong inhibitory effect at lower concentrations.

[0134] The present invention has been described in detail above. It will be apparent to those skilled in the art that the present invention may be implemented in a wide range under equivalent parameters, concentrations and conditions without departing from the spirit and scope of the present invention and without the need for unnecessary experimentation. Although the present invention provides specific embodiments, it should be understood that further improvements may be made to the present invention. In short, according to the principles of the present invention, this application intends to include any changes, uses or improvements to the present invention, including changes made by conventional techniques known in the art that depart from the scope disclosed in this application. Applications of some of the basic features may be made within the scope of the following appended claims.

Claims

1. Lactobacillus plantarum, characterized in that The plant lactobacillus is plant lactobacillus (Lactobacillus plantarum) DZ041, and its deposit number in the General Microbiological Center of China National Microbiological Culture Collection Committee is CGMCC No. 20835.

2. A bacterial agent, characterized in that: The bacterial agent contains the Lactobacillus plantarum according to claim 1.

3. A fermentation broth, characterized in that The Lactobacillus plantarum fermentation liquid is obtained by fermenting the Lactobacillus plantarum according to claim 1.

4. A composition, characterized in that The composition comprises the plant lactobacillus according to claim 1 and / or the bacterial agent according to claim 2 and / or the fermentation broth according to claim 3.

5. The application of the plant lactobacillus according to claim 1 in any of the following: A1) Use in inhibiting or slowing the growth of fungi or bacteria and / or use in preparing products for inhibiting and / or slowing the growth of fungi or bacteria; A2) Use in the lysis of fungi and / or in the preparation of lysis fungal products; A3) Use in destroying the integrity of fungal cell membranes and / or use in preparing products that destroy the integrity of fungal cell membranes; A4) Preparation for use in treating or assisting in treating diseases caused by fungi; A5) Preparation for use in treating or assisting in treating diseases caused by fungi.

6. Use of the plant lactobacillus according to claim 1 in inhibiting bacterial biofilm formation or preparing a product for inhibiting bacterial biofilm formation.

7. Use of the bacterial agent according to claim 2 in any of the following: A1) Use in inhibiting or slowing the growth of fungi and / or use in preparing products for inhibiting and / or slowing the growth of fungi; A2) Use in the lysis of fungi and / or in the preparation of lysis fungal products; A3) Use in destroying the integrity of fungal cell membranes and / or use in preparing products that destroy the integrity of fungal cell membranes; A4) Preparation for use in treating or assisting in treating diseases caused by fungi; A5) Preparation for use in treating or assisting in treating diseases caused by fungi; A6) Use in inhibiting bacterial biofilm formation or in preparing products for inhibiting bacterial biofilm formation.

8. Use of the bacterial agent according to claim 2 in inhibiting bacterial biofilm formation or in preparing a product for inhibiting bacterial biofilm formation.

9. Use of the fermentation broth according to claim 3 in inhibiting or slowing down the formation of fungal biofilms, in preparing products for inhibiting or slowing down the formation of fungal biofilms, and / or in preparing products for treating or assisting in treating diseases caused by fungi.

10. The use according to any one of claims 5 to 9, characterized in that The fungi are Malassezia furfur, Pseudomonas aeruginosa and / or Staphylococcus aureus.

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