KR20200135145A - Lactobacillus paracasei wikim0110 having antibacterial activity against clostridioides difficile and composition comprising the same - Google Patents

Abstract

The present invention relates to Lactobacillus paracasei WiKim0110 and a composition including the same. The Lactobacillus paracasei WiKim0110 according to the present invention is a probiotic having antibacterial activity against harmful pathogens (Clostridioides difficile), and can be used in various ways for purposes such as prevention, improvement, and treatment of harmful pathogens infectious diseases of humans or animals.

Description

[LACTOBACILLUS PARACASEI WIKIM0110 HAVING ANTIBACTERIAL ACTIVITY AGAINST CLOSTRIDIOIDES DIFFICILE AND COMPOSITION COMPRISING THE SAME}

The present invention relates to a Lactobacillus paracasei WiKim0110 strain and a composition comprising the same.

Clostridioides difficile infection (CDI) disease is an infectious disease caused by Clostridioides difficile , a major intestinal pathogenic bacterium well known worldwide. Worldwide, there are 453,000 infections per year, and in the United States, there are about 15,000 direct deaths from Clostridioides difficile. Approximately 124,000 and 18,005 people are infected annually in Europe and the UK. Patients diagnosed with CDI in Asia, especially East Asia, have a high prevalence, as in Europe and North America. CDI is usually accompanied by abdominal pain, diarrhea and fever. In addition, CDI can cause a variety of symptoms, including diarrhea and gastrointestinal colitis that occur in the intestine after extensive antibiotic use and can be life-threatening.

In addition, Clostridioides difficile is an opportunistic pathogen that resides in the intestine, but does not cause any symptoms in people with strong intestinal immunity, and is observed at a high rate in the intestine of newborns within 1 month of life. If antibiotics continue to be taken to treat diarrhea or Pseudomembranous colitis, the number of Clostridioides difficile, currently antibiotic-resistant, increases rapidly in the intestine.Severe severe diarrhea due to toxins secreted by these bacteria As a result, toxicity and tolerance problems arise, and development of a new CDI treatment technology is required (Non-Patent Document 1).

Therefore, the importance of improvement, prevention or treatment of infectious diseases caused by Clostridioides difficile is on the rise. Thus, in the present invention, a study was conducted on lactic acid bacteria that effectively improves the infectious disease caused by Clostridioides difficile.

Fecal transplant and fecal bank. Science and Technology Policy Volume 216, 14~15, (2016)

Accordingly, the present inventors tried to find a strain that exhibits antibacterial activity against Clostridioides difficile , as a result of which, as a result of isolating and identifying Lactobacillus paracasei having antibacterial activity against Clostridioides difficile The present invention has been completed.

Accordingly, an object of the present invention is to provide an antimicrobial composition for Clostridioides difficile comprising Lactobacillus paracasei WiKim0110, a culture thereof, a lysate thereof, an extract thereof, or a bacteriocin produced therefrom as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of infectious diseases caused by Clostridioides difficile comprising Lactobacillus paracasei WiKim0110, a culture thereof, a lysate thereof, or an extract thereof as an active ingredient.

In addition, another object of the present invention is to provide a food composition for preventing or improving infectious diseases caused by Clostridioides difficile comprising Lactobacillus paracasei WiKim0110, a culture thereof, a lysate thereof, or an extract thereof as an active ingredient.

In addition, another object of the present invention is to provide a fermentation starter comprising Lactobacillus paracasei WiKim0110, its culture, its lysate or its extract as an active ingredient.

In addition, another object of the present invention is to provide a feed additive or feed composition comprising Lactobacillus paracasei WiKim0110, its culture, its lysate or its extract as an active ingredient.

Another object of the present invention is to provide a probiotic composition comprising Lactobacillus paracasei WiKim0110, a culture thereof, a lysate thereof, or an extract thereof as an active ingredient.

In order to achieve the object of the present invention as described above, the present invention provides a Lactobacillus paracasei WiKim0110 ( Lactobacillus paracasei WiKim0110).

The Lactobacillus paracasei WiKim0110 according to the present invention is a Lactobacillus paracasei strain derived from infant feces. Although the Lactobacillus paracasei WiKim0110 in the present invention was isolated and identified from the feces of infants, the route of obtaining it is not limited thereto.

It was confirmed that the lactobacillus strain derived from infant feces isolated through the examples of the present invention is excellent as probiotics and exhibits excellent antibacterial effects against harmful pathogens. In addition, 16S rRNA gene sequencing analysis for identification and classification of microorganisms showed that it has the nucleic acid sequence of SEQ ID NO: 1. In addition, as a result of analysis, the microorganism showed the highest molecular phylogenetic relationship with the subspecies Lactobacillus paracasei strains.

Accordingly, the microorganism of the present invention having the 16S rRNA gene sequence of SEQ ID NO: 1 was named Lactobacillus paracasei CBA3611, and was deposited with the National Academy of Agricultural Sciences on March 22, 2019 (accession number KACC81092BP). In the present invention, the Lactobacillus paracasei CBA3611 ( Lactobacillus paracasei CBA3611) can be used interchangeably with the Lactobacillus paracasei WiKim0110 ( Lactobacillus paracasei WiKim0110).

The strain of Lactobacillus paracasei WiKim0110 (Accession No. KACC81092BP) of the present invention is a Gram-positive bacterium, is a facultative anaerobe that can grow in anaerobic and aerobic conditions, and takes the form of a bacillus.

In the present invention, "probiotics" are understood to mean "live microorganisms that have a beneficial effect on the health of the host by improving the intestinal microbial environment of the host in the gastrointestinal tract of animals including humans". Probiotics are living microorganisms with probiotic activity, and when fed to humans or animals in the form of single or multiple strains in the form of dried cells or fermented products, they can have a beneficial effect on the intestinal flora of the host.

In the examples of the present invention, it was confirmed that the Lactobacillus paracasei WiKim0110 of the present invention exhibits high antibacterial activity in an antibacterial test against harmful pathogens. In addition, through sequence analysis of Lactobacillus paracasei WiKim0110, it was confirmed that the WiKim0110 has a bacteriocin, enterolysin A gene. The Lactobacillus paracasei WiKim0110 according to the present invention is a probiotic and can be used in various ways for antibacterial of harmful pathogens in humans or animals.

Accordingly, one embodiment of the present invention provides a composition comprising Lactobacillus paracasei WiKim0110, a culture thereof, a lysate thereof, or an extract thereof as an active ingredient.

Lactobacillus paracasei WiKim0110 included in the composition according to the present invention may exist as dead or live cells, and may also exist in a dried or lyophilized form. Forms of lactic acid bacteria suitable for inclusion in various compositions and methods of formulation are well known to those skilled in the art.

The present invention also provides a composition for antibacterial comprising Lactobacillus paracasei WiKim0110, a culture thereof, a lysate thereof, an extract thereof, or a bacteriocin produced therefrom.

In one embodiment, the antimicrobial composition may be one having an antimicrobial activity against Clostridioides difficile. The Clostridioides difficile is a representative harmful pathogen causing diarrhea, and diarrhea by producing intestinal toxin [toxin A], cytotoxin [toxin B], and binary toxin And can cause an inflammatory reaction.

Accordingly, the present invention provides a composition for preventing, improving or treating infectious diseases caused by harmful pathogens, including Lactobacillus paracasei WiKim0110, a culture thereof, a lysate thereof, or an extract thereof.

Lactobacillus paracasei WiKim0110 according to the present invention may be included in medicines, health functional foods, foods, feeds, feed additives or fermentation starters due to this advantageous action.

Accordingly, the present invention provides a pharmaceutical composition for the prevention or treatment of infectious diseases caused by Clostridioides difficile comprising Lactobacillus paracasei WiKim0110, a culture thereof, a lysate thereof, or an extract thereof as an active ingredient. In the present invention, the infectious disease may be diarrhea, toxic giant colon, perforation, sepsis, or pseudomembranous colitis, but is not limited thereto.

When the composition of the present invention is used as a pharmaceutical composition, the pharmaceutical composition of the present invention may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant is an excipient, boron Releases, sweeteners, binders, coatings, expanding agents, lubricants, lubricants or flavoring agents may be used.

The pharmaceutical composition may be preferably formulated as a pharmaceutical composition, including at least one pharmaceutically acceptable carrier in addition to the active ingredients described above for administration.

For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders are, but are not limited to, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, lacquercanth or sodium oleate, sodium stearate, magnesium stearate, sodium benzo Eate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthan gum, and the like. As acceptable pharmaceutical carriers for compositions formulated as liquid solutions, sterilization and biocompatible ones, saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added if necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare injectable formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets.

Further, it may be preferably formulated according to each disease or ingredient using a method disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA) as an appropriate method in the field.

In addition, the present invention may be a food composition for preventing or improving infectious diseases caused by Clostridioides difficile comprising Lactobacillus paracasei WiKim0110, its culture, its lysate or its extract as an active ingredient.

The food composition according to the present invention may be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods. Foods containing the composition of the present invention include, for example, health functional foods, beverages, vitamin complexes, and health supplement foods.

The food composition of the present invention may include ingredients that are commonly added during food production, and include, for example, proteins, carbohydrates, fats, nutrients, flavoring agents and flavoring agents. Examples of the aforementioned carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)] and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is made of drinks and beverages, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts may be additionally included.

The present invention provides a composition having antibacterial activity against Clostridioides difficile comprising Lactobacillus paracasei WiKim0110, a culture thereof, a lysate thereof, or an extract thereof as an active ingredient.

In one embodiment, the composition may be a composition for oral administration comprising the Lactobacillus paracasei WiKim0110 strain present as live bacteria.

Lactobacillus paracasei WiKim0110 according to the present invention may be included in medicine, health functional food, food, feed, feed additive or fermentation starter due to this advantageous action.

In addition, the food composition containing the strain as an active ingredient may include a beverage such as fermented milk.

Accordingly, the present invention provides a fermentation starter consisting of Lactobacillus paracasei WiKim0110, a culture thereof, a lysate thereof, or an extract thereof.

In addition, the present invention can be used as a feed additive or feed containing Lactobacillus paracasei WiKim0110, a culture thereof, a lysate thereof, or an extract thereof as an active ingredient.

When used as a feed additive, the composition may be a high concentration of 20 to 90% or may be prepared in the form of powder or granules. The feed additives are organic acids such as citric acid, humic acid, adipic acid, lactic acid, malic acid, or phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate, and polyphosphate (polyphosphate), polyphenol, catechin, alpha-tocopherol, rosemary Any one or more of natural antioxidants such as extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, and phytic acid may be further included. When used as feed, the composition may be formulated in a conventional feed form, and may include a common feed component.

The feed additives and feed include grains, such as crushed or crushed wheat, oats, barley, corn and rice; Vegetable protein feeds such as feeds based on rape, soybeans, and sunflowers; Animal protein feeds such as blood meal, meat meal, bone meal and fish meal; It may further include a dry component composed of sugar and dairy products, for example, various milk powders and whey powder, and may further include nutritional supplements, digestion and absorption enhancers, growth accelerators, and the like.

The feed additive may be administered to the animal alone or may be administered in combination with other feed additives in an edible carrier. In addition, the feed additives may be easily administered to animals as top dressings, directly mixed with animal feeds, or in an oral formulation separate from feed. When the feed additive is administered separately from animal feed, it can be prepared in an immediate release or sustained release formulation by combining it with a pharmaceutically acceptable edible carrier as well known in the art. Such edible carriers may be solid or liquid, such as corn starch, lactose, sucrose, soy flakes, peanut oil, olive oil, sesame oil and propylene glycol. When a solid carrier is used, the feed additive may be a tablet, a capsule, a powder, a troche or a sugar-containing tablet, or a top dressing in a microdispersible form. When a liquid carrier is used, the feed additive may be a gelatin soft capsule, or a formulation of a syrup, suspension, emulsion, or solution.

In addition, the feed additive and feed may contain adjuvants, such as preservatives, stabilizers, wetting or emulsifying agents, solution accelerators, and the like. The feed additive may be used by dipping, spraying, or mixing to add to animal feed.

The feed additive or feed of the present invention can be applied to a number of animal diets, including mammals, poultry and fish.

As the mammal, pigs, cows, sheep, goats, experimental rodents, and experimental rodents, as well as pets (eg, dogs, cats), etc. can be used, and as the poultry, chickens, turkeys, ducks, geese, pheasants, And it may be used for quail and the like, and may be used for trout as the fish, but is not limited thereto.

The amount of the Lactobacillus paracasei WiKim0110 strain contained in the composition according to the present invention may be about 10 ⁶ to 10 ¹² cfu/g, for example, 10 ⁷ to 10 ¹¹ cfu/g, 10 ⁸ to 10 ¹⁰ cfu. May be /g. In the case of administering the strain, it is preferable to administer it in a viable state, and it may be killed before ingestion or administered in an attenuation state. In addition, in the case of manufacturing using a culture supernatant or the like, a sterilization process through a heat treatment process may be additionally performed. The amount of strain and the degree of daily intake required to have the minimum efficacy may vary depending on the body or health condition of the ingestor, but may generally be about 10 ⁶ to 10 ¹² cfu/g, such as 10 ⁷ to 10 ¹¹ cfu/g , 10 ⁸ to 10 ¹⁰ cfu/g.

Advantages and features of the present invention, and a method of achieving them will become apparent with reference to embodiments described below in detail. However, the present invention is not limited to the embodiments disclosed below, but will be implemented in a variety of different forms, and only the present embodiments are intended to complete the disclosure of the present invention, and the general knowledge in the technical field to which the present invention belongs It is provided to completely inform the scope of the invention to those who have it, and the invention is only defined by the scope of the claims.

Since Lactobacillus paracasei WiKim0110 according to the present invention exhibits excellent antibacterial activity against harmful pathogens (Clostridioides difficile), it is used as a probiotic for preventing, improving or treating infectious diseases caused by harmful pathogens of humans or animals. It can be used in various ways for Furthermore, it can be usefully used as a fermentation starter.

Figure 1 shows the molecular phylogenetic analysis results of the Lactobacillus paracasei WiKim0110 ( Lactobacillus paracasei WiKim0110) strain using the 16S rRNA gene sequence.
Figure 2 is a result of confirming the antibacterial activity of the strain of Lactobacillus paracasei WiKim0110 ( Lactobacillus paracasei WiKim0110) against Clostridioides difficile JCM 1296. (A) Control group, (B) Lactobacillus paracasei WiKim0110 culture solution treatment group.

Hereinafter, the present invention will be described in detail through examples. The following examples are merely illustrative of the present invention, and the scope of the present invention is not limited to the following examples.

[Example]

Example 1: Isolation and identification of Lactobacillus paracasei strain

Infant fecal samples were suspended in PBS (Phosphate-buffered saline), spread on MRS solid medium, and incubated at 30° C. for 2 days. A single colony of bacteria obtained after cultivation was collected by loop and cultured in MRS broth. DNA extraction was performed using the QIAamp DNA Mini Kit (QIAgen, Germany). The extracted DNA was confirmed using a 1% agarose gel, and using the genomic DNA extracted to amplify the 16S rRNA gene as a template, 27F(5'-AGAGTTTGATCCTGGCTCAG-3'), 1492R(5'-CTACGGCTACCTTGTTACGA-3' ) PCR was performed using a primer, and PCR conditions were performed 30 cycles with denaturation 95°C for 1 minute, annealing 45°C for 1 minute, and extension 72°C for 1 minute and 30 seconds. The obtained PCR product was sequenced using a nucleotide sequence analyzer. Bacterial identification was performed through a method of searching for bacteria with the highest similarity to the nucleotide sequence of the 16S rRNA gene of the strain using the EzBioCloud (https://www.ezbiocloud.net/identify) database.

The strain isolated through the embodiment of the present invention is a result of nucleotide sequence analysis of the 16S rRNA gene for the identification of microorganisms, the strain is Lactobacillus paracasei subsp. Tolerans DSM 20258 strain and 100% homology was confirmed (FIG. 1 ), but the Ortho ANI (Average Nucleotide Identity) value comparing the entire genome with the corresponding strain was 98.3%, which was confirmed as another strain with a difference in the genome.

Accordingly, the microorganism of the present invention was named Lactobacillus paracasei CBA3611, and was deposited with the National Academy of Agricultural Sciences on March 22, 2019 (accession number KACC81092BP).

Example 2: Confirmation of antibacterial activity of Lactobacillus paracasei WiKim0110

A single colony of Lactobacillus paracasei WiKim0110 isolated through Example 1 was inoculated in MRS liquid medium and cultured in an incubator at 30° C. for 48 hours. The strain and the supernatant were separated by centrifugation, and the supernatant was prepared with a strain culture solution from which bacteria were completely removed using a 0.2 μl pore size filter. The culture solution was concentrated 5 times using a vacuum concentrator.

To confirm the antimicrobial activity of the Lactobacillus paracasei WiKim0110, Clostridioides difficile JCM 1296 was prepared as a harmful pathogen. The harmful pathogens were smeared on a CDC medium (BD BBL™ CDC Anaerobe 5%, Sheep Blood Agar-Becton Dickinson), and a paper disc was inoculated on the medium smeared with harmful pathogens. 90 µl of the concentrated Lactobacillus paracasei WiKim0110 culture solution was absorbed into a paper disc, and after 24 hours, the culture of harmful pathogens and the culture inhibition by Lactobacillus paracasei WiKim0110 were confirmed (FIG. 2 ). At this time, Figure 2 (A) is the result of the control group not treated with the culture medium of Lactobacillus paracasei WiKim0110, (B) is the result of the experimental group treated with the culture medium of Lactobacillus paracasei WiKim0110.

As a result, as shown in Figure 2, it was confirmed that the effect of inhibiting the culture of Clostridioides difficile appears in the paper disc of the group treated with the Lactobacillus paracasei WiKim0110 culture solution.

As a result, it was confirmed that it has the ability to inhibit growth against Clostridioides difficile (FIG. 2).

Example 3: Analysis of the bacteriocin genome of Lactobacillus paracasei WiKim0110

For the genome analysis of the Lactobacillus paracasei WiKim0110, the extracted DNA was sequenced using PacBio RS II (Pacific Biosciences), and the gene was analyzed.

Particularly, as a result of searching through the BAGEL4 webserver (http://bagel4.molgenrug.nl), whether or not the genome of Lactobacillus paracasei WiKim0110 is bacteriocin, the enterorlysin A gene corresponding to class III was identified. Enterolysin A was identified as a bacteriocin with a broad spectrum of growth inhibition that bursts and kills cells while cutting N-acetylmuramoyl and L-amino acid residues on the cell wall.

<SEQ ID NO: 2> Amino acid sequence of enterolysin A

MSYTNKAGANGSSVANRYHDVGASGARANAAYKNNAAAYTAVVGDGGVYVGGYVWGAGTVANANSVGHTSDTKKDYAVYARDMAARYGTSDAGGAGTGKSHWVTHWGDHTDYGYARWGTKKAADANGTTTVDASKSAAASARAVTRNVNVAYGHGRWDVTNGSGDNGAGMNYHDYKVDHGSVKYRVHTVSGWWVAKGDRNDTVNGCAGNAGVDGVTAGSYKAYYRSTTRAGWGVVCDDGTSYTDTYAGGDRGSSN

Institute of Agricultural Biotechnology KACC81092BP 20190322

<110> Korea Food Research Institute <120> LACTOBACILLUS PARACASEI WIKIM0110 HAVING ANTIBACTERIAL ACTIVITY AGAINST CLOSTRIDIOIDES DIFFICILE AND COMPOSITION COMPRISING THE SAME <130> P19R10C0364 <160> 4 <170> KoPatentIn 3.0 <210> 1 <211> 1570 <212> RNA <213> Unknown <220> <223> Lactobacillus sp. Wikim0110 <220> <221> gene <222> (1)..(1570) <400> 1 ttatatgaga gtttgatcct ggctcaggat gaacgctggc ggcgtgccta atacatgcaa 60 gtcgaacgag ttctcgttga tgatcggtgc ttgcaccgag attcaacatg gaacgagtgg 120 cggacgggtg agtaacacgt gggtaacctg cccttaagtg ggggataaca tttggaaaca 180 gatgctaata ccgcatagat ccaagaaccg catggttctt ggctgaaaga tggcgtaagc 240 tatcgctttt ggatggaccc gcggcgtatt agctagttgg tgaggtaatg gctcaccaag 300 gcgatgatac gtagccgaac tgagaggttg atcggccaca ttgggactga gacacggccc 360 aaactcctac gggaggcagc agtagggaat cttccacaat ggacgcaagt ctgatggagc 420 aacgccgcgt gagtgaagaa ggctttcggg tcgtaaaact ctgttgttgg agaagaatgg 480 tcggcagagt aactgttgtc ggcgtgacgg tatccaacca gaaagccacg gctaactacg 540 tgccagcagc cgcggtaata cgtaggtggc aagcgttatc cggatttatt gggcgtaaag 600 cgagcgcagg cggtttttta agtctgatgt gaaagccctc ggcttaaccg aggaagcgca 660 tcggaaactg ggaaacttga gtgcagaaga ggacagtgga actccatgtg tagcggtgaa 720 atgcgtagat atatggaaga acaccagtgg cgaaggcggc tgtctggtct gtaactgacg 780 ctgaggctcg aaagcatggg tagcgaacag gattagatac cctggtagtc catgccgtaa 840 acgatgaatg ctaggtgttg gagggtttcc gcccttcagt gccgcagcta acgcattaag 900 cattccgcct ggggagtacg accgcaaggt tgaaactcaa aggaattgac gggggcccgc 960 acaagcggtg gagcatgtgg tttaattcga agcaacgcga agaaccttac caggtcttga 1020 catcttttga tcacctgaga gatcaggttt ccccttcggg ggcaaaatga caggtggtgc 1080 atggttgtcg tcagctcgtg tcgtgagatg ttgggttaag tcccgcaacg agcgcaaccc 1140 ttatgactag ttgccagcat ttagttgggc actctagtaa gactgccggt gacaaaccgg 1200 aggaaggtgg ggatgacgtc aaatcatcat gccccttatg acctgggcta cacacgtgct 1260 acaatggatg gtacaacgag ttgcgagacc gcgaggtcaa gctaatctct taaagccatt 1320 ctcagttcgg actgtaggct gcaactcgcc tacacgaagt cggaatcgct agtaatcgcg 1380 gatcagcacg ccgcggtgaa tacgttcccg ggccttgtac acaccgcccg tcacaccatg 1440 agagtttgta acacccgaag ccggtggcgt aaccctttta gggagcgagc cgtctaaggt 1500 gggacaaatg attagggtga agtcgtaaca aggtagccgt aggagaacct gcggctggat 1560 cacctccttt 1570 <210> 2 <211> 255 <212> PRT <213> Unknown <220> <223> Enterolysin A <400> 2 Met Ser Tyr Thr Asn Lys Ala Gly Ala Asn Gly Ser Ser Val Ala Asn 1 5 10 15 Arg Tyr His Asp Val Gly Ala Ser Gly Ala Arg Ala Asn Ala Ala Tyr 20 25 30 Lys Asn Asn Ala Ala Ala Tyr Thr Ala Val Val Gly Asp Gly Gly Val 35 40 45 Tyr Val Gly Gly Tyr Val Trp Gly Ala Gly Thr Val Ala Asn Ala Asn 50 55 60 Ser Val Gly His Thr Ser Asp Thr Lys Lys Asp Tyr Ala Val Tyr Ala 65 70 75 80 Arg Asp Met Ala Ala Arg Tyr Gly Thr Ser Asp Ala Gly Gly Ala Gly 85 90 95 Thr Gly Lys Ser His Trp Val Thr His Trp Gly Asp His Thr Asp Tyr 100 105 110 Gly Tyr Ala Arg Trp Gly Thr Lys Lys Ala Ala Asp Ala Asn Gly Thr 115 120 125 Thr Thr Val Asp Ala Ser Lys Ser Ala Ala Ala Ser Ala Arg Ala Val 130 135 140 Thr Arg Asn Val Asn Val Ala Tyr Gly His Gly Arg Trp Asp Val Thr 145 150 155 160 Asn Gly Ser Gly Asp Asn Gly Ala Gly Met Asn Tyr His Asp Tyr Lys 165 170 175 Val Asp His Gly Ser Val Lys Tyr Arg Val His Thr Val Ser Gly Trp 180 185 190 Trp Val Ala Lys Gly Asp Arg Asn Asp Thr Val Asn Gly Cys Ala Gly 195 200 205 Asn Ala Gly Val Asp Gly Val Thr Ala Gly Ser Tyr Lys Ala Tyr Tyr 210 215 220 Arg Ser Thr Thr Arg Ala Gly Trp Gly Val Val Cys Asp Asp Gly Thr 225 230 235 240 Ser Tyr Thr Asp Thr Tyr Ala Gly Gly Asp Arg Gly Ser Ser Asn 245 250 255 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Forward primer <220> <221> gene <222> (1)..(20) <223> 27F <400> 3 agagtttgat cctggctcag 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer <220> <221> gene <222> (1)..(20) <223> 1492R <400> 4 ctacggctac cttgttacga 20

Claims (9)

Antimicrobial against Clostridioides difficile containing Lactobacillus paracasei WiKim0110 (accession number KACC81092BP), its culture, its lysate, its extract, or bacteriocin produced therefrom as an active ingredient Composition. Pharmaceuticals for the prevention or treatment of infectious diseases caused by Clostridioides difficile containing Lactobacillus paracasei WiKim0110 (accession number KACC81092BP), its culture, its lysate or its extract as an active ingredient Composition. The method of claim 2,
The infectious disease caused by Clostridioides difficile is a pharmaceutical composition that is caused by diarrhea, toxic giant colon, perforation, sepsis or pseudomembranous colitis. Food for the prevention or improvement of infectious diseases caused by Clostridioides difficile containing Lactobacillus paracasei WiKim0110 (accession number KACC81092BP), its culture, its lysate or its extract as an active ingredient Composition. The method of claim 4,
The infectious disease caused by the Clostridioides difficile is diarrhea, toxic giant colon, perforation, sepsis or gastrointestinal colitis. The method of claim 4,
The food is a health functional food, food composition. Fermentation starter comprising Lactobacillus paracasei WiKim0110 ( Lactobacillus paracasei WiKim0110; accession number KACC81092BP), its culture, its lysate or its extract as an active ingredient. A feed additive or feed composition comprising Lactobacillus paracasei WiKim0110 ( Lactobacillus paracasei WiKim0110; accession number KACC81092BP), its culture, its lysate or its extract as an active ingredient. A probiotic composition comprising Lactobacillus paracasei WiKim0110 ( Lactobacillus paracasei WiKim0110; accession number KACC81092BP), a culture thereof, a lysate thereof, or an extract thereof as an active ingredient.

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