WO2023229309A1 - Souche de lactobacillus paracasei uos1 et composition antibactérienne la comprenant - Google Patents

Souche de lactobacillus paracasei uos1 et composition antibactérienne la comprenant Download PDF

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WO2023229309A1
WO2023229309A1 PCT/KR2023/006893 KR2023006893W WO2023229309A1 WO 2023229309 A1 WO2023229309 A1 WO 2023229309A1 KR 2023006893 W KR2023006893 W KR 2023006893W WO 2023229309 A1 WO2023229309 A1 WO 2023229309A1
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strain
uos1
antibacterial
lactobacillus paracasei
antibacterial composition
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PCT/KR2023/006893
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English (en)
Korean (ko)
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기호삼
백준석
윤성건
최용준
정선욱
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주식회사 라비오
서울시립대학교 산학협력단
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Publication of WO2023229309A1 publication Critical patent/WO2023229309A1/fr

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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P1/00—Disinfectants; Antimicrobial compounds or mixtures thereof
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P3/00—Fungicides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus

Definitions

  • the present invention relates to Lactobacillus paracasei UOS1 strain having antibacterial activity and its use.
  • bacteria such as Listeria monocytogenes, Salmonella genus, Staphylococcus aureus and some pathogenic Escherichia coli and fungi such as Candida albicans are opportunistic pathogens that pose a risk to human health. It can be threatening.
  • Cosmetics are made up of high molecular lipids or water as the main ingredient and a mixture of various other substances. Among these substances, they contain glycerin and sorbitol, which serve as carbon sources for microorganisms, and many contain amino acid derivatives and proteins as nitrogen sources. Bacteria and molds that use carbon and nitrogen sources as mediators can easily multiply.
  • preservatives must be used to protect cosmetics for a long period of time. Since cosmetics are repeatedly used on the skin, there is a need to develop preservatives that have excellent biosafety and less skin irritation.
  • Parabens a preservative widely used in cosmetics, drugs, and processed foods, have been used for a long time because they have a wide spectrum of antibacterial activity against Gram-positive and Gram-negative bacteria and are effective.
  • synthetic raw materials have excellent antibacterial activity, but they can cause dermatitis, allergies, and abnormal hormone secretion, and there is a risk that they may remain on the skin without being biodegraded. Therefore, they are a safe and excellent antibacterial alternative to synthetic raw materials. There is a need to discover materials and commercialize them.
  • Natural preservatives to replace synthetic preservatives include plant extracts such as safflower extract, grapefruit extract, hinokitol, and magnolol, but these are difficult to be widely used due to their high production cost, and their low preservative efficacy allows microorganisms to proliferate. There is a risk that you can do it.
  • Metabolites with antibacterial activity produced by microorganisms include biosurfactant and bacteriocin (Jiang et al., 2012; Sharma and Saharan, 2014).
  • biosurfactant and bacteriocin there are various enzymes and siderophores that are antibacterial and antifungal substances produced by microorganisms. reported (Nagarajkumar et al., 2004).
  • the present inventors found that they can produce metabolites that inhibit the growth of microorganisms, have antibacterial activity against various types of microorganisms, and are stable even in a wide range of pH. It was confirmed that it can exhibit antibacterial activity and the present invention was completed.
  • the present invention is intended to solve the problems of the prior art described above, and the purpose of the present invention is to provide a natural antibacterial agent derived from a new strain with excellent biosafety and antibacterial activity that can replace conventional synthetic preservatives.
  • Lactobacillus paracasei UOS1 strain deposited with accession number KACC 81213BP is provided.
  • the strain has the ability to produce citric acid, lactic acid, and acetic acid, and produces the citric acid, lactic acid, and acetic acid in a range of 35 to 40:15 to 20:44. It can be produced at a weight ratio of 46.
  • an antibacterial composition comprising the above-mentioned strain, its culture, lysate, extract or purified product as an active ingredient is provided.
  • the strain may be cultured by adding natural products or natural extracts.
  • the antibacterial composition is Escherichia coli , Staphylococcus aureus , Pseudomonas aeruginosa , Candida albicans , and Aspergillus brazil. It may have antibacterial activity against one or more microorganisms selected from the group consisting of Aspergillus brasiliensis .
  • the antibacterial composition may have antibacterial activity in the pH range of 4.0 to 9.0.
  • a cosmetic composition containing the antibacterial composition is provided.
  • an external skin composition comprising the antibacterial composition is provided.
  • a method for producing an antibacterial composition is provided, including the step.
  • the metabolites of the strain can exhibit antibacterial activity against various types of microorganisms and can therefore be added to medicines, foods and cosmetics and used as preservatives to increase the shelf life of the products.
  • the culture medium containing the metabolites of the above strain exhibits stable antibacterial activity even in a wide pH range and can be widely applied to various types of products.
  • the antibacterial composition has low toxicity to the human body and excellent antibacterial effect, so it can replace conventional synthetic preservatives.
  • Figure 1 is a 16s rDNA base sequence result for Lactobacillus paracasei USO1 strain according to an embodiment of the present invention.
  • Figure 2 shows the results of component analysis of Lactobacillus paracasei UOS1 strain culture medium according to an embodiment of the present invention.
  • Figures 3 and 4 show the results of evaluating the antibacterial activity of Lactobacillus paracasei UOS1 strain culture medium according to an embodiment of the present invention.
  • the numerical range includes the values defined in the range above. Every maximum numerical limit given throughout this specification includes all lower numerical limits as if the lower numerical limit were explicitly written out. Every minimum numerical limit given throughout this specification includes every higher numerical limit as if such higher numerical limit was clearly written. All numerical limits given throughout this specification will include all better numerical ranges within the broader numerical range, as if the narrower numerical limits were clearly written.
  • Lactobacillus paracasei UOS1 strain deposited with accession number KACC 81213BP is provided.
  • the “Lactobacillus paracasei UOS1 strain” is a new lactic acid bacterium with 98.11% 16s rDNA base sequence homology to Lactobacillus paracasei NRIC 1938 strain.
  • the present inventors isolated the above-mentioned strains from cabbage kimchi aged for a long time, and among them, the Lactobacillus paracasei UOS1 strain was isolated, which has antibacterial activity against various types of microorganisms and can stably exhibit antibacterial activity even in a wide range of pH. did.
  • the strain produces organic acids such as citric acid, lactic acid, and acetic acid as metabolites, effectively inhibiting microbial growth in cosmetics and foods, and also producing various substances that inhibit the growth of microorganisms. Through this, the growth of contamination-causing bacteria, pathogenic bacteria, and putrefactive bacteria can be suppressed.
  • the Lactobacillus paracasei UOS1 strain can maximize antibacterial activity by producing citric acid, lactic acid, and acetic acid as metabolites at an optimal ratio.
  • the Lactobacillus paracasei UOS1 strain can produce citric acid, lactic acid, and acetic acid at a weight ratio of 35 to 40:15 to 20:44 to 46, and optimal antibacterial activity can be achieved within this range. .
  • the present inventors cultivated the strain under optimal conditions through media optimization technology, analyzed metabolites resulting from the culture, and confirmed that the culture medium can effectively inhibit the growth of various types of microorganisms.
  • an antibacterial composition comprising the above-mentioned strain, its culture, lysate, extract or purified product as an active ingredient is provided.
  • Metabolites of the strain can exhibit antibacterial activity against various types of microorganisms, and the strain or its culture solution can be usefully used as a natural antibacterial agent.
  • the antibacterial composition includes the strain or its culture, lysate, extract, or purified product, and contains microbial inhibitory metabolites such as organic acids, so it can exhibit excellent antibacterial activity.
  • the culture may be the culture itself obtained by cultivating the strain, a culture supernatant obtained by removing the strain therefrom, or a concentrate or freeze-dried product of the culture supernatant.
  • the strain may be cultured by adding natural products or natural extracts.
  • the strain can be cultured under various conditions or media, and when cultured by adding a natural product or a natural extract consisting of its active ingredients in the culture step, the antibacterial activity can be enhanced due to additionally produced or changed metabolites.
  • the type of the natural product or natural extract is not particularly limited, but is preferably an extract of a fruit or plant containing sugar that the strain can use as energy, such as sugarcane extract, persimmon extract, or apple extract,
  • One or more may be selected from the group consisting of blueberry extract, licorice extract, raspberry extract, prickly pear extract, rice extract, agar extract, blackberry extract, corn extract, pear extract, banana extract, peach extract, and blueberry extract.
  • the culture includes the steps of (a) pre-culturing the Lactobacillus paracasei UOS1 strain deposited under the accession number KACC 81213BP; (b) inoculating the strain into a culture medium and fermenting it; and (c) removing the strain and recovering the supernatant.
  • the culture is prepared by adding the strain to one or more carbon sources selected from the group consisting of glucose, lactose, fructose, mannose, galactose, xylose, ribose, maltose, sucrose, dextrin, glycerin, and inulin; and soy protein hydrolyzate, skim milk powder, potassium nitrate, ammonium nitrate, ammonium sulfate, ammonium oxalate, and diammonium inulin phosphate.
  • carbon sources selected from the group consisting of glucose, lactose, fructose, mannose, galactose, xylose, ribose, maltose, sucrose, dextrin, glycerin, and inulin
  • soy protein hydrolyzate skim milk powder, potassium nitrate, ammonium nitrate, ammonium sulfate, ammonium oxalate, and diammonium inul
  • the carbon source or nitrogen source used in the culture medium is, for example, about 0.1 to 30 w/v%, 1 to 25 w/v%, 1 to 20 w/v%, 1 to 10 w/v%, 1 to 5 w/v. %, or 1 to 1.5 w/v%.
  • the culture medium may contain a commonly used salt, and the salt may include, for example, one or more salts selected from the group consisting of potassium nitrate, potassium sulfate, magnesium carbonate, magnesium sulfate, and magnesium nitrate,
  • the salt may be included, for example, at about 0.001 to 10 w/v%, 0.1 to 7 w/v%, or 0.5 to 5 w/v%.
  • the culture medium may include about 0.01 to 10 w/v%, 0.05 to 5 w/v%, or 0.1 to 2 w/v% of yeast autolysate, and the culture medium has a pH of about 2.0 to 7.0. It may be, but is not limited to this.
  • the culture is performed by culturing the strain under temperature conditions of, for example, about 20 to 42°C, about 25 to 35°C, or about 27 to 30°C for, for example, about 24 to 144 hours, 48 to 120 hours, or 15 to 96 hours. It may have been obtained by doing so.
  • the culture conditions of the strain may include the step of adding the strain to the medium and fermenting it, and the fermentation step may use a commonly used fermentation method, for example, the method may use fed batch fermentation. You can.
  • the extract may be obtained by extracting the isolated culture of the strain with various organic solvents.
  • the lysate is obtained by isolating the strain from the culture of the strain, disrupting the cell wall or cell membrane through mechanical methods or nonmechanical methods, and releasing intracellular products. It may be.
  • the purified product may be obtained by cultivating the strain and completing the fermentation process, or by extracting and purifying the active ingredient therefrom.
  • the antibacterial composition can exhibit antibacterial activity against various types of microorganisms, such as Escherichia coli, Staphylococcus aureus , Pseudomonas aeruginosa , and Candida albicans ( It may have antibacterial activity against one or more microorganisms selected from the group consisting of Candida albicans ) and Aspergillus brasiliensis , but is not limited thereto.
  • microorganisms such as Escherichia coli, Staphylococcus aureus , Pseudomonas aeruginosa , and Candida albicans ( It may have antibacterial activity against one or more microorganisms selected from the group consisting of Candida albicans ) and Aspergillus brasiliensis , but is not limited thereto.
  • the antibacterial composition may have antibacterial activity in the pH range of 4.0 to 9.0.
  • the antibacterial composition can maintain its antibacterial activity stably despite a wide range of pH changes, it can be easily applied to various products such as food and external skin preparations as well as cosmetic products formulated in various forms.
  • a cosmetic composition or external skin preparation containing the antibacterial composition is provided.
  • the skin external preparation or cosmetic composition can be formulated by conventional methods.
  • information disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA can be referred to, and in formulating cosmetic compositions, International cosmetic ingredient dictionary, 6th ed., The cosmetic, Toiletry and Fragrance. Association, Inc., Washington, 1995.
  • the cosmetic composition can be prepared in the form of a general emulsified formulation and solubilized formulation.
  • lotion such as softening lotion or nourishing lotion
  • Emulsions such as facial lotion and body lotion
  • Creams such as nutritional cream, moisture cream, eye cream, etc.; essence; cosmetic ointment; spray; gel; pack; sunscreen; makeup base; Foundation, such as liquid type, solid type, or spray type; powder
  • Make-up removers such as cleansing creams, cleansing lotions, and cleansing oils
  • it may be formulated as a cleansing agent such as cleansing foam, soap, or body wash, but is not limited to this.
  • the skin external preparation may be formulated as an ointment, patch, gel, cream, or spray, but is not limited thereto.
  • other ingredients may be appropriately mixed in each formulation of the skin external preparation or cosmetic composition within the range that does not impair the purpose according to the present invention, depending on the type of formulation or purpose of use.
  • the skin external preparation or cosmetic composition may contain a commonly acceptable carrier, such as oil, water, surfactant, moisturizer, lower alcohol, thickener, chelating agent, colorant, preservative, fragrance, etc., may be appropriately mixed. It is not limited.
  • the acceptable carrier may vary depending on the formulation. For example, when formulated into ointments, pastes, creams or gels, animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or these are used as carrier ingredients. Mixtures may be used.
  • lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, or a mixture thereof may be used as a carrier ingredient, and in the case of a spray, chlorochloride It may further include propellants such as fluorohydrocarbon, propane, butane or dimethyl ether.
  • a solvent, solubilizing agent, or emulsifying agent may be used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl benzoate, Propylene glycol, 1,3-butyl glycol oil may be used, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol aliphatic esters, polyethylene glycol or fatty acid esters of sorbitan. You can.
  • the carrier ingredient includes water, a liquid diluent such as ethanol or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester.
  • a liquid diluent such as ethanol or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester.
  • Suspending agents, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used.
  • alkali metal salts of fatty acids fatty acid hemiester salts, fatty acid protein hydrolyzates, isethionates, lanolin derivatives, fatty alcohols, vegetable oils, glycerol, sugars, etc. are used as carrier ingredients.
  • alkali metal salts of fatty acids fatty acid hemiester salts, fatty acid protein hydrolyzates, isethionates, lanolin derivatives, fatty alcohols, vegetable oils, glycerol, sugars, etc.
  • fatty acids fatty acid hemiester salts
  • fatty acid protein hydrolyzates isethionates
  • lanolin derivatives fatty alcohols
  • vegetable oils glycerol, sugars, etc.
  • the skin external preparation or cosmetic composition may contain fatty substances, organic solvents, solubilizers, thickeners, gelling agents, softeners, antioxidants, suspending agents, stabilizers, and foaming agents commonly used in the industry, depending on the quality or function of the final product. ), fragrances, surfactants, water, ionic or non-ionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic activators, common in cosmetics. It may additionally contain auxiliaries commonly used in the fields of cosmetology or dermatology, such as any other ingredients used.
  • auxiliaries and their mixing ratio may be appropriately selected so as not to affect the desirable properties of the cosmetic composition according to the present invention.
  • MRS solid medium 1% peptone, 1% beef. extract, 0.4% yeast extract, 2% glucose, 0.5% sodium acetate monohydrate, 0.1% tween 80, 0.2% monobasic potassium phosphate, 0.2% triammonium citrate, 0.02% magnesium sulfate, 0.005% manganese sulfate monohydrate, 1.5% agar.
  • a single colony formed after culturing was taken, inoculated into the same medium using a stroke plate method, and then cultured at 30°C for 3 days to obtain a pure single species.
  • the isolated single strain was inoculated into 3 mL of MRS liquid medium and cultured with shaking at 30 °C for 2 days.
  • the cultured bacterial fluid was mixed 1:1 (v/v) with 80% glycerol solution and placed in an ultra-low temperature freezer (-80 °C). It was stored, re-inoculated as needed, and used in future experiments.
  • 16s rDNA nucleotide sequence analysis was performed as follows.
  • the culture medium was incubated at 13,000 rpm and 4°C for 3 minutes. Cells were recovered by centrifugation.
  • gDNA Chromosomal DNA
  • the obtained PCR product was confirmed by electrophoresis on a 1% agarose gel at a voltage of 120 V for 20 minutes, and a DNA amplification product of about 1.5 kb in length was cut and purified using a DNA fragment purification kit (Intron lifetechnology).
  • the isolated strain as a new plant-derived lactic acid bacteria, was named Lactobacillus paracasei UOS1 and deposited with the Korea Agricultural Microorganism Conservation Center (KACC) under accession number 81213BP on May 10, 2022.
  • KACC Korea Agricultural Microorganism Conservation Center
  • Example 1 Strain culture and fermentation purification
  • the identified microorganism produces substances that inhibit microbial growth, such as organic acids, through fermentation and secretes them outside the cell. Therefore, the microorganism was cultured and a culture medium was obtained therefrom.
  • Appropriate culture medium conditions for the identified microorganism include 1 to 1.5 w/v% of glucose or lactose, 1 to 7 w/v% of skim milk or soy protein hydrolyzate, and 0.2 to 0.4 w/v of yeast autolyzate yeast extract. v%, malt extract at 0.001 to 0.4 w/v%, and each inorganic salt at 0.001 to 1.5 w/v%, and was sterilized at 121°C for 15 minutes before use.
  • the prepared culture medium was cultured at 25 to 35°C for 48 to 96 hours at 200 rpm, then subjected to ultrasonic extraction for 1 to 3 hours to obtain an extract, and the supernatant was separated at 12,000 rpm for 15 to 30 minutes and filtered through a 0.22 ⁇ m membrane filter. It was recovered after sterilization.
  • the recovered and filtered culture solution was purified using an impurity remover.
  • Types of impurity removers include activated carbon, ion exchange resin, white clay, magnesium silicate, diatomaceous earth, tangerine peel, and lemon peel.
  • the amount of impurity remover used was 0.1% to 10% of the recovered culture medium, stirred at 25 to 35°C for 20 to 90 minutes, and then recovered after sterilization using a 0.22 ⁇ m membrane filter.
  • a fermented purified product was obtained in the same manner as in Example 1, but the culture was prepared using a culture medium mixed with 1 to 10 w/v% of natural extract (sugar cane extract) when culturing the identified microorganism.
  • a commercially available Lactobacillus paracasei single strain was inoculated into MRS liquid medium sterilized at 121°C for 15 minutes, cultured at 25 to 35°C for 48 to 96 hours at 200 rpm, and centrifuged at 12,000 rpm for 15 to 30 minutes. The separated supernatant was sterilized using a 0.22 ⁇ m membrane filter and then recovered.
  • the recovered culture medium was recovered in the same manner as in the example using an impurity remover.
  • Purified water was added and mixed at a 1:1 ratio to the culture, and a sample solution for analysis was prepared using a 0.2 ⁇ m membrane filter.
  • the components contained in the mixed solution were analyzed using an HPLC (Waters Alliance HPLC 2695, USA) system.
  • the column used was MetaCarb 87H (250 ⁇ 4.6 mm, Agilent).
  • the detector used was a PDA (Photodiode Array Detector).
  • the highest concentration of the substance was detected in the three peak retention times excluding the extraction solvent peak, and this was detected through the selective ion monitoring mode and the library of compounds included in the Agilent MassHunter workstation software. As a result of the analysis, the detected peaks were confirmed to be citric acid, lactic acid, and acetic acid.
  • citric acid was analyzed as 0.7%, lactic acid as 0.3%, and acetic acid as 0.8%.
  • citric acid was analyzed at the level of 0.05 to 0.2%, lactic acid at the level of 0.6 to 1.5%, and acetic acid at the level of 0.3 to 0.6%, so the proportion of organic acids was different, resulting in differences in antibacterial activity. can be estimated.
  • the minimum inhibitory concentration (MIC) and minimum killing concentration (MBC) of the strain cultures obtained in the examples were measured.
  • the culture concentration of the strain obtained in the example was diluted using a 2-fold dilution method using sterilized water to obtain a volume of 100 ⁇ L per well in a 96 well plate.
  • TSB medium Tryptic Soy Broth
  • C. albicans a culture obtained by sterilizing sterilized distilled water with 4% glucose and 1% peptone was cultured in an incubator at 30°C with shaking for 48 hours.
  • A. brasiliensis a culture obtained by sterilizing commercially available PDB medium (P otato Dextrose Broth ) from difco and shaking it in an incubator at 30°C for 48 hours was used.
  • PDB medium P otato Dextrose Broth
  • the suspension method was prepared by diluting with the optimal medium used for each strain.
  • the 96-well plate with completed inoculation was incubated for 24 or 48 hours at the optimal growth temperature for each strain, then the turbidity was checked, and the culture concentration in the well where turbidity was not visible was set to MIC, the minimum concentration that inhibits bacterial growth. decided.
  • MIC concentration 1.5% of agar was added to the optimal growth medium for each strain, and 20 ⁇ L of each was smeared on a plate solidified in a petri dish. After culturing for 24 or 48 hours at the optimal growth temperature for each strain, no bacterial growth was observed. The minimum concentration was determined as MBC.
  • Example 1 E. coli ⁇ 1.0% ⁇ 1.0% S. aureus ⁇ 1.0% ⁇ 1.0% P. aeruginosa ⁇ 1.0% ⁇ 1.0% C. albicans ⁇ 1.0% ⁇ 1.0% A. brasiliensis 2.0% 2.0%
  • Example 2 E. coli ⁇ 1.0% ⁇ 1.0% S. aureus ⁇ 1.0% ⁇ 1.0% P. aeruginosa ⁇ 1.0% ⁇ 1.0% C. albicans ⁇ 0.5% ⁇ 0.5% A. brasiliensis ⁇ 2.5% ⁇ 2.5% Comparative Example 1 E. coli 15% 15% S. aureus 25% 25% P. aeruginosa 15% 15% C. albicans 30% 40% A.
  • brasiliensis 50% >50% Comparative Example 2 E. coli 15% 15% S. aureus 20% 25% P. aeruginosa 15% 15% C. albicans 40% 40% A. brasiliensis 40% 50% Comparative Example 3 E. coli 20% 15% S. aureus 20% 25% P. aeruginosa 20% 20% C. albicans 40% 50% A. brasiliensis >50% >50%
  • Example 1 showed strong antibacterial activity with MIC and MBC results of 1.0% or less for harmful bacteria E. coli, S. aureus, P. aeruginosa, and C. albicans , and A. brasiliensis . At a concentration of about 2.0%, the growth of microorganisms was effectively inhibited.
  • Example 2 cultured with the natural extract showed antibacterial activity equivalent to that of Example 1, and its antibacterial activity against C. albicans was evaluated at a higher level.

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Abstract

La présente invention concerne une souche Lactobacillus paracasei UOS1 présentant une activité antibactérienne et une composition antibactérienne comprenant une culture, un lysat, un extrait ou un filtrat de celle-ci en tant que principe actif.
PCT/KR2023/006893 2022-05-26 2023-05-22 Souche de lactobacillus paracasei uos1 et composition antibactérienne la comprenant WO2023229309A1 (fr)

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KR1020220064542A KR102450693B1 (ko) 2022-05-26 2022-05-26 락토바실러스 파라카제이 uos1 균주, 및 이를 포함하는 항균 조성물

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KR102450693B1 (ko) * 2022-05-26 2022-10-06 주식회사 라비오 락토바실러스 파라카제이 uos1 균주, 및 이를 포함하는 항균 조성물

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US20100226892A1 (en) * 2009-03-04 2010-09-09 L'oreal Use of probiotic microorganisms to limit skin irritation
KR20130118625A (ko) * 2012-04-20 2013-10-30 조선대학교산학협력단 항진균 활성을 갖는 락토바실러스 플란타룸 및 이를 포함하는 항진균 조성물
WO2015120098A1 (fr) * 2014-02-04 2015-08-13 Micro-Nature Llc Systèmes, procédés et compositions relatifs aux combiomiques
KR102134820B1 (ko) * 2019-06-05 2020-07-17 서울시립대학교 산학협력단 항균 활성을 갖는 락토바실러스 파라카세이 아종 톨러란스 sw1 균주 및 이의 용도
KR102450693B1 (ko) * 2022-05-26 2022-10-06 주식회사 라비오 락토바실러스 파라카제이 uos1 균주, 및 이를 포함하는 항균 조성물

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KR101990820B1 (ko) 2017-05-22 2019-06-19 전남대학교산학협력단 균주 배양액의 항균 활성 예측을 위한 정보 제공 방법 및 균주 배양액을 포함하는 항균용 조성물

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