KR102207512B1 - Lactobacillus jensenii swpm104 which has antibacterial activity - Google Patents
Lactobacillus jensenii swpm104 which has antibacterial activity Download PDFInfo
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- KR102207512B1 KR102207512B1 KR1020200069283A KR20200069283A KR102207512B1 KR 102207512 B1 KR102207512 B1 KR 102207512B1 KR 1020200069283 A KR1020200069283 A KR 1020200069283A KR 20200069283 A KR20200069283 A KR 20200069283A KR 102207512 B1 KR102207512 B1 KR 102207512B1
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- swpm104
- lactobacillus
- genseni
- composition
- lactobacillus genseni
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Abstract
Description
본 발명은 항균 활성을 갖는 락토바실러스 젠세니 SWPM104 및 이를 이용한 항균 조성물, 약학적 조성물 및 식품 조성물에 대한 것이다.The present invention relates to Lactobacillus genseni SWPM104 having antibacterial activity, and to an antibacterial composition, a pharmaceutical composition, and a food composition using the same.
락토바실러스 속 (Lactobacillus sp.) 미생물은 동형 또는 이형발효를 하는 젖산 간균으로서 사람을 포함한 동물의 장관(Intestinal tract) 및 유제품이나 채소의 발효과정에서 흔히 볼 수 유산균의 하나이다. 락토바실러스 속 미생물은 장내 pH를 산성으로 유지시켜, 대장균(E. coli)이나 클로스트리디움(Clostridium)과 같은 유해균의 번식을 억제하고 설사와 변비를 개선할 뿐만 아니라, 비타민 합성, 항암 작용, 혈청 콜레스테롤 저하 등의 역할을 하는 것으로 알려져 있다. Lactobacillus sp. is one of the lactic acid bacillus commonly found in the intestinal tract of animals including humans and in the fermentation process of dairy products or vegetables as a lactic acid bacillus that undergo homogeneous or heterogeneous fermentation. Lactobacillus microorganisms keep the intestinal pH acidic, inhibiting the reproduction of harmful bacteria such as E. coli and Clostridium, improving diarrhea and constipation, as well as synthesizing vitamins, anti-cancer action, and serum. It is known to play a role such as lowering cholesterol.
최근에는 락토바실러스를 이용한 식품 또는 약학적 조성물들이 개발되고 있는데, 예컨대, 한국등록특허 10-1884326호는 락토바실러스 브레비스 G1(Lactobacillus brevis G1) 균주(KCCM11898P)을 포함하는 식품 알레르기 예방 및 치료용 조성물을 개시하고 있고, 한국등록특허 10-1873898호는 락토바실러스 람노서스 CJLR1505(Lactobacillus rhamnosus CJLR1505)(KCCM11721P)의 사균체를 이용한 동물 사료용 조성물을 개시하고 있다.Recently, food or pharmaceutical compositions using Lactobacillus have been developed.For example, Korean Patent No. 10-1884326 discloses a composition for preventing and treating food allergies including Lactobacillus brevis G1 strain (KCCM11898P). Korean Patent Registration No. 10-1873898 discloses a composition for animal feed using dead cells of Lactobacillus rhamnosus CJLR1505 (Lactobacillus rhamnosus CJLR1505) (KCCM11721P).
본 발명자들은 항균 활성을 갖는 락토바실러스 속 균주에 대하여 연구하던 중 한국 임산부들의 질내 시료에서 채취한 신규 락토바실러스 젠세니(Lactobacillus jensenii) SWPM104의 항균 활성을 확인하고 본 발명을 완성하였다.The present inventors confirmed the antimicrobial activity of the novel Lactobacillus jensenii SWPM104 obtained from vaginal samples of Korean pregnant women while studying the strain of the genus Lactobacillus having antibacterial activity and completed the present invention.
본 발명의 목적은 신규 락토바실러스 젠세니 균주를 제공하는 것이다.An object of the present invention is to provide a novel Lactobacillus genseni strain.
상기 목적을 달성하기 위하여 본 발명은 항균 활성을 갖는 락토바실러스 젠세니(Lactobacillus jensenii) SWPM104 (기탁번호 : KCTC 14168BP) 및 이를 이용한 항균 조성물, 약학적 조성물 및 식품 조성물을 제공한다.In order to achieve the above object, the present invention provides Lactobacillus jensenii) SWPM104 (accession number: KCTC 14168BP) having antibacterial activity and an antibacterial composition, pharmaceutical composition, and food composition using the same.
본 발명의 락토바실러스 젠세니 SWPM104는 항균 활성, 내담즙성, 내산성을 가지며, 항균 조성물, 약학적 조성물 및 식품 조성물에 이용 가능하다.Lactobacillus genseni SWPM104 of the present invention has antibacterial activity, bile resistance, acid resistance, and can be used in antibacterial compositions, pharmaceutical compositions and food compositions.
도 1는 락토바실러스 젠세니 SWPM104 균주가 0.3 중량% 담즙산염 농도에서 증식한 것을 보여준다.
도 2은 락토바실러스 젠세니 SWPM104 균주의 항생제 감수성 평가 결과이다.
도 3는 락토바실러스 젠세니 SWPM104 균주의 과산화수소 생성력을 확인한 결과를 보여준다.1 shows that the Lactobacillus genseni SWPM104 strain was grown at a concentration of 0.3% by weight bile salt.
2 is a result of evaluation of antibiotic susceptibility of Lactobacillus genseni SWPM104 strain.
Figure 3 shows the results of confirming the hydrogen peroxide production ability of Lactobacillus genseni SWPM104 strain.
본 발명은,The present invention,
항균 활성을 갖는 락토바실러스 젠세니(Lactobacillus jensenii) SWPM104 (기탁번호 : KCTC 14168BP)에 대한 것이다.It is for Lactobacillus jensenii SWPM104 (accession number: KCTC 14168BP) having antibacterial activity.
또한 본 발명은 락토바실러스 젠세니 SWPM104, 그 배양물 또는 그 추출물을 포함하는 항균 조성물에 대한 것이다.In addition, the present invention relates to an antimicrobial composition comprising Lactobacillus genseni SWPM104, a culture thereof or an extract thereof.
또한 본 발명은 락토바실러스 젠세니 SWPM104, 그 배양물 또는 그 추출물을 포함하는, 세균성 질염, 아토피성 피부염, 녹농균 감염으로 인한 호흡기 질환, 녹농균 감염으로 인한 폐렴, 치아우식증, 충치, 장염, 및 식중독으로 구성되는 군으로부터 선택되는 하나 이상의 질병의 예방 또는 치료용 약학적 조성물에 대한 것이다.In addition, the present invention relates to bacterial vaginosis, atopic dermatitis, respiratory disease due to Pseudomonas aeruginosa infection, pneumonia due to Pseudomonas aeruginosa infection, dental caries, tooth decay, enteritis, and food poisoning, including Lactobacillus genseni SWPM104, its culture or its extract. It relates to a pharmaceutical composition for the prevention or treatment of one or more diseases selected from the group consisting of.
또한 본 발명은 락토바실러스 젠세니 SWPM104, 그 배양물 또는 그 추출물을 포함하는, 세균성 질염, 아토피성 피부염, 녹농균 감염으로 인한 호흡기 질환, 녹농균 감염으로 인한 폐렴 치아우식증, 충치, 장염, 및 식중독으로 구성되는 군으로부터 선택되는 하나 이상의 질병의 예방 또는 개선용 식품 조성물에 대한 것이다.In addition, the present invention comprises Lactobacillus genseni SWPM104, a culture thereof or an extract thereof, bacterial vaginosis, atopic dermatitis, respiratory disease due to Pseudomonas aeruginosa infection, pneumonia due to Pseudomonas aeruginosa infection, dental caries, tooth decay, enteritis, and food poisoning. It relates to a food composition for preventing or improving one or more diseases selected from the group.
이하, 본 발명을 자세히 설명한다.Hereinafter, the present invention will be described in detail.
락토바실러스 젠세니(Lactobacillus Jenseni ( Lactobacillus jenseniiLactobacillus jensenii ) SWPM104) SWPM104
본 발명의 락토바실러스 젠세니(Lactobacillus jensenii) SWPM104 (기탁번호 : KCTC 14168BP)는 항균 활성을 갖는다. 바람직하게는 본 발명의 락토바실러스 젠세니 SWPM104은 질염 유발균, 충치 유발균, 아토피성 피부염 유발균, 녹농균, 장염균 및 식중독균으로 구성되는 군으로부터 선택되는 하나 이상에 대하여 항균 활성을 갖는다. 더욱 바람직하게는 본 발명의 락토바실러스 젠세니 SWPM104은 가르드네렐라 바기날리스(Gardnerella vaginalis), 아토포비움 바기날리스(Atopobium vaginalis), 스트렙토쿠스 뮤탄스(Streptococcus mutans), 녹농균(Pseudomonas aeruginosa), 표피포도구균(Staphylococcus epidermidis), 바실러스 세레우스(Bacillus cereus) 및 살모넬라 엔테리티디스(Salmonella enteritidis)으로 구성되는 군으로부터 선택되는 하나 이상에 대하여 항균 활성을 갖는다. Lactobacillus jensenii of the present invention ( Lactobacillus jensenii ) SWPM104 (accession number: KCTC 14168BP) has antibacterial activity. Preferably, the Lactobacillus genseni SWPM104 of the present invention has antimicrobial activity against at least one selected from the group consisting of vaginitis-causing bacteria, tooth decay-inducing bacteria, atopic dermatitis-inducing bacteria, Pseudomonas aeruginosa, enteritis and food poisoning bacteria. More preferably, the Lactobacillus genseni SWPM104 of the present invention is Gardnerella vaginalis, Atopobium vaginalis, Streptococcus mutans, Pseudomonas aeruginosa, It has antimicrobial activity against at least one selected from the group consisting of Staphylococcus epidermidis, Bacillus cereus, and Salmonella enteritidis.
본 발명의 락토바실러스 젠세니 SWPM104은 pH3 또는 pH 7의 조건 하 증식능을 갖는다. 또한 본 발명의 락토바실러스 젠세니 SWPM104은 0.3 중량% 담즙산을 포함하는 배지 조건 하 증식능을 갖는다. 이때, 증식능을 갖는다는 것은 상기 pH 조건 하 또는 담즙산 농도 조건 하, 3시간 배양 시 배양 후 생균 수가 증가하는 것을 의미한다. Lactobacillus genseni SWPM104 of the present invention has a proliferative ability under the conditions of pH3 or pH7. In addition, Lactobacillus genseni SWPM104 of the present invention has a proliferative ability under medium conditions containing 0.3% by weight bile acid. At this time, having a proliferative ability means that the number of viable cells increases after incubation for 3 hours under the above pH condition or under the bile acid concentration condition.
본 발명의 락토바실러스 젠세니 SWPM104은 세균성 질염을 유발하는 세균에 대한 항균 활성과 질내 부착능이 우수하다.The Lactobacillus genseni SWPM104 of the present invention is excellent in antibacterial activity and vaginal adhesion against bacteria that cause bacterial vaginosis.
락토바실러스 젠세니 SWPM104은 질 내의 우점종(dominant species)이고 과산화수소(H2O2)를 생산하는 능력이 우수하며, 질 상피세포에 대한 우수한 접착능력과 질염 유발 세균에 대해 우수한 생육 억제 활성을 나타낸다. 그러므로 본 발명은 락토바실러스 젠세니 SWPM104 균주 또는 이의 배양물을 유효성분으로 포함하는 부인과 질환의 예방 및 치료를 위한 건강 보조 식품, 의약품(약학적 조성물 등) 및 의약외품, 화장품을 제공한다. Lactobacillus genseni SWPM104 is a dominant species in the vagina, has excellent ability to produce hydrogen peroxide (H2O2), and exhibits excellent adhesion to vaginal epithelial cells and excellent growth inhibitory activity against yeast infection-causing bacteria. Therefore, the present invention provides health supplements, pharmaceuticals (pharmaceutical compositions, etc.), quasi-drugs, and cosmetics for the prevention and treatment of gynecological diseases comprising Lactobacillus genseni SWPM104 strain or a culture thereof as an active ingredient.
또한 락토바실러스 젠세니 SWPM104은 충치 유발 세균에 대해 우수한 항균 활성을 나타내기 때문에 본 발명은 락토바실러스 젠세니 SWPM104 균주 또는 이의 배양물을 유효성분으로 충치 예방 및 개선을 위한 건강 보조 식품, 일반 식품, 의약품(약학적 조성물 등), 의약외품을 제공한다. In addition, since Lactobacillus genseni SWPM104 exhibits excellent antibacterial activity against tooth decay-causing bacteria, the present invention uses Lactobacillus genseni SWPM104 strain or a culture thereof as an active ingredient as a health supplement for preventing and improving caries, general foods, and pharmaceuticals. (Pharmaceutical composition, etc.), provides quasi-drugs.
또한 락토바실러스 젠세니 SWPM104은 녹농균에 대해 우수한 항균 활성을 나타낸다. 그러므로 본 발명은 락토바실러스 젠세니 SWPM104 균주 또는 이의 배양물을 유효성분으로 녹농균 감염증의 예방 및 치료를 위한 의약품 등을 제공한다.In addition, Lactobacillus genseni SWPM104 exhibits excellent antibacterial activity against Pseudomonas aeruginosa. Therefore, the present invention provides a drug for the prevention and treatment of Pseudomonas aeruginosa infection as an active ingredient, Lactobacillus genseni SWPM104 strain or a culture thereof.
그러나 락토바실러스 젠세니 SWPM104은 대장균(Escherichia coli)에 대하여는 항균 활성이 없다.However, Lactobacillus genseni SWPM104 has no antibacterial activity against Escherichia coli.
항균 조성물Antibacterial composition
본 발명은 락토바실러스 젠세니 SWPM104, 그 배양물 또는 그 추출물을 포함하는 항균 조성물에 대한 것이다. 바람직하게는 상기 항균 조성물은 가르드네렐라 바기날리스(Gardnerella vaginalis), 아토포비움 바기날리스(Atopobium vaginalis), 스트렙토쿠스 뮤탄스(Streptococcus mutans), 녹농균(Pseudomonas aeruginosa), 표피포도구균(Staphylococcus epidermidis), 바실러스 세레우스(Bacillus cereus) 및 살모넬라 엔테리티디스(Salmonella enteritidis)로 구성되는 군으로부터 선택되는 하나 이상의 세균에 대하여 항균 활성을 갖는다. The present invention relates to an antimicrobial composition comprising Lactobacillus genseni SWPM104, a culture thereof or an extract thereof. Preferably, the antibacterial composition is Gardnerella vaginalis, Atopobium vaginalis, Streptococcus mutans, Pseudomonas aeruginosa, and Staphylococcus epidermidis ), Bacillus cereus (Bacillus cereus) and Salmonella enteritidis (Salmonella enteritidis) has an antimicrobial activity against one or more bacteria selected from the group consisting of.
“항균”은 특정 농도에서 미생물의 성장 또는 생존을 감소, 방지, 억제, 또는 제거하는 능력을 의미한다. 항균용 조성물이란 항미생물제를 총칭하는 의미인 항생제와 같은 의미일 수 있고, 항균제, 살균제, 방부제, 보존제 또는 제균제와 같은 의미일 수 있으며, 바람직하게는 그람양성균 및 그람음성균으로 이루어진 군에서 선택된 1 종 이상의 미생물의 발육과 생활 기능을 저지 또는 억제할 수 있는 물질 즉, 항세균 효력이 있는 물질을 의미한다.“Antibacterial” means the ability to reduce, prevent, inhibit, or eliminate the growth or survival of a microorganism at a specific concentration. The antimicrobial composition may have the same meaning as antibiotics, which is a generic term for antimicrobial agents, and may have the same meaning as antimicrobial agents, fungicides, preservatives, preservatives or disinfectants, preferably 1 selected from the group consisting of Gram-positive bacteria and Gram-negative bacteria. It refers to a substance capable of inhibiting or inhibiting the development and life function of more than one species, that is, a substance having antibacterial effect.
본 발명의 항균 조성물은, 질염에 대하여 사용하는 경우 질 내외에 직접 투여하거나 적절한 형태의 조성물로 제조하여 투여할 수 있다. 상기와 같은 측면에서, 상기 항균 조성물은, 건강 보조 식품, 의약품, 의약외품, 화장품 형태일 수 있다. 본 발명의 항균 조성물은, 충치 예방 또는 치료의 경우는 입 안에서 씹거나 씹어서 삼키거나 가글할 수 있는 형태의 조성물로 제조하여 투여할 수 있다. 상기와 같은 측면에서, 상기 조성물은, 건강 보조 식품, 일반 식품, 의약품, 의약외품의 형태일 수 있다. 본 발명의 항균 조성물은, 아토피성 피부염의 예방, 개선 또는 치료용 조성물일 수 있다. 상기와 같은 측면에서, 상기 조성물은, 의약품, 의약외품, 화장품의 형태일 수 있다When the antibacterial composition of the present invention is used against vaginitis, it may be administered directly into or out of the vagina, or may be prepared and administered in an appropriate form. In the above aspect, the antibacterial composition may be in the form of health supplements, pharmaceuticals, quasi-drugs, and cosmetics. The antimicrobial composition of the present invention may be prepared and administered in a form capable of being chewed or chewed in the mouth, swallowed or gargled in the case of preventing or treating caries. In the above aspect, the composition may be in the form of health supplements, general foods, pharmaceuticals, and quasi-drugs. The antibacterial composition of the present invention may be a composition for preventing, improving or treating atopic dermatitis. In the above aspect, the composition may be in the form of pharmaceuticals, quasi-drugs, and cosmetics.
약학적 조성물Pharmaceutical composition
본 발명은 락토바실러스 젠세니 SWPM104, 그 배양물 또는 그 추출물을 포함하는 약학적 조성물에 대한 것이다. 바람직하게는 본 발명은 락토바실러스 젠세니 SWPM104, 그 배양물 또는 그 추출물을 포함하는, 세균성 질염, 아토피성 피부염, 녹농균 감염으로 인한 질병 및 충치로 구성되는 군으로부터 선택되는 하나 이상의 질병의 예방 또는 치료용 약학적 조성물에 대한 것이다. 녹농균에 의한 감염성 질병은 호흡기 질환, 패혈증, 전신감염, 기도감염, 낭포성 섬유증, 녹농균 모낭염, 골수염, 괴저성 농창(ecthyma gangraenosum), 방광염, 중이염, 농흉(empyema), 폐렴, 뇌막염, 복막염, 각막염, 내안구염 및 균혈증을 포함하나 이에 본 발명이 제한되는 것은 아니고, 녹농균에 의하여 발생할 수 있는 이차감염을 모두 포함한다.The present invention relates to a pharmaceutical composition comprising Lactobacillus genseni SWPM104, a culture thereof or an extract thereof. Preferably, the present invention is the prevention or treatment of one or more diseases selected from the group consisting of bacterial vaginosis, atopic dermatitis, diseases caused by Pseudomonas aeruginosa infection and tooth decay, including Lactobacillus genseni SWPM104, a culture thereof or an extract thereof. It is about a pharmaceutical composition for use. Infectious diseases caused by Pseudomonas aeruginosa are respiratory diseases, sepsis, systemic infections, airway infections, cystic fibrosis, Pseudomonas aeruginosa folliculitis, osteomyelitis, ecthyma gangraenosum, cystitis, otitis media, empyema, pneumonia, meningitis, peritonitis, keratitis. , Including endophthalmitis and bacteremia, but the present invention is not limited thereto, and all secondary infections that may be caused by Pseudomonas aeruginosa are included.
본 발명의 항균용 조성물의 투여량은 투여 대상에서 상기 유해 세균을 사멸시키는 효과를 발휘하기 위해 요구되는 정도의 양을 의미하는데, 제형의 종류 및 투여 대상의 연령, 체중, 일반 건강 상태, 성별 및 식이, 투여 경로, 투여 시간 및 기간을 비롯한 다양한 인자에 따라 조절될 수 있다. 락토바실러스 젠세니 SWPM104 균주, 그 파쇄물, 그 배양액 또는 그 추출물은, 조성물 총 중량을 기준으로, 0.01중량% 내지 10중량%로 포함될 수 있다. 일 구체예에서, 상기 락토바실러스 젠세니 SWPM104 균주, 그 파쇄물, 그 배양액 또는 그 추출물은, 조성물 총 중량을 기준으로, 0.1중량%~5중량%로 포함될 수 있다. 상기와 같은 측면에서, 락토바실러스 젠세니 SWPM104 균주, 그 파쇄물, 그 배양액 또는 그 추출물은, 조성물 총 중량을 기준으로, 0.01중량% 이하로도 포함될 수 있다.The dosage of the antimicrobial composition of the present invention refers to the amount required to exert the effect of killing the harmful bacteria in the subject to be administered, the type of the dosage form and the age, weight, general health condition, sex, and It can be adjusted according to various factors including diet, route of administration, time and duration of administration. Lactobacillus genseni SWPM104 strain, its lysate, its culture solution or its extract may be contained in an amount of 0.01% to 10% by weight based on the total weight of the composition. In one embodiment, the Lactobacillus genseni SWPM104 strain, its lysate, its culture solution or its extract may be included in an amount of 0.1% to 5% by weight, based on the total weight of the composition. In the above aspect, the Lactobacillus genseni SWPM104 strain, its lysate, its culture solution or its extract may be included in an amount of 0.01% by weight or less based on the total weight of the composition.
상기 약학적 조성물은 제조에 통상적으로 사용하는 적절한 담체, 부형제 및 희석제를 더 포함할 수 있다. 상기 약학적 조성물은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다. 본 발명에 따른 상기 약학조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유 등을 포함한 다양한 화합물 혹은 혼합물을 들 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 추출물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트(calcium carbonate), 수크로스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로젤라틴 등이 사용될 수 있다.The pharmaceutical composition may further include a suitable carrier, excipient, and diluent commonly used in preparation. The pharmaceutical compositions may be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. . Carriers, excipients and diluents that can be included in the pharmaceutical composition according to the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, Various compounds or mixtures including calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. . In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and these solid preparations include at least one excipient in the extract such as starch, calcium carbonate, sucrose. ) Or lactose (lactose), gelatin, etc. are mixed to prepare. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, solvents, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients, such as wetting agents, sweetening agents, fragrances, and preservatives, may be included. . Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogelatin, and the like may be used.
식품 조성물Food composition
본 발명은 락토바실러스 젠세니 SWPM104, 그 배양물 또는 그 추출물을 포함하는 식품 조성물에 대한 것이다. 바람직하게는 본 발명은 락토바실러스 젠세니 SWPM104, 그 배양물 또는 그 추출물을 포함하는, 세균성 질염, 아토피성 피부염, 녹농균 감염으로 인한 질병 및 충치로 구성되는 군으로부터 선택되는 하나 이상의 질병의 예방 또는 개선용 식품 조성물에 대한 것이다. 녹농균에 의한 감염성 질병은 호흡기 질환, 패혈증, 전신감염, 기도감염, 낭포성 섬유증, 녹농균 모낭염, 골수염, 괴저성 농창(ecthyma gangraenosum), 방광염, 중이염, 농흉(empyema), 폐렴, 뇌막염, 복막염, 각막염, 내안구염 및 균혈증을 포함하나 이에 본 발명이 제한되는 것은 아니고, 녹농균에 의하여 발생할 수 있는 이차감염을 모두 포함한다. The present invention relates to a food composition comprising Lactobacillus genseni SWPM104, a culture thereof or an extract thereof. Preferably, the present invention is to prevent or improve one or more diseases selected from the group consisting of bacterial vaginosis, atopic dermatitis, diseases caused by Pseudomonas aeruginosa infection and tooth decay, including Lactobacillus genseni SWPM104, a culture or extract thereof For food composition. Infectious diseases caused by Pseudomonas aeruginosa are respiratory diseases, sepsis, systemic infections, airway infections, cystic fibrosis, Pseudomonas aeruginosa folliculitis, osteomyelitis, ecthyma gangraenosum, cystitis, otitis media, empyema, pneumonia, meningitis, peritonitis, keratitis. , Including endophthalmitis and bacteremia, but the present invention is not limited thereto, and all secondary infections that may be caused by Pseudomonas aeruginosa are included.
상기 식품 조성물은 건강 식품 조성물일 수 있으며, 식품 조성물의 제형은 특별히 한정되지 않으나, 예를 들어, 정제, 환제, 연질 및 경질 캅셀제, 과립제, 드링크제, 캐러멜, 분말 등으로 제형화 될 수 있다. 각 제형의 식품 조성물은 유효 성분 이외에 해당 분야에서 통상적으로 사용되는 성분들을 제형 또는 사용 목적에 따라 당업자가 어려움 없이 적의 선정하여 배합할 수 있으며, 다른 원료와 동시에 적용할 경우 상승 효과가 일어날 수 있다. 상기 유효 성분의 투여량 결정은 당업자의 수준 내에 있으며, 이의 1일 투여 용량은 예를 들어 락토바실러스 젠세니가 105 내지 1013 CFU/일, 보다 구체적으로는 108 내지 1010 CFU/일이 될 수 있으나, 이에 제한되지 않으며, 투여하고자 하는 대상의 연령, 건강 상태, 합병증 등의 다양한 요인에 따라 달라질 수 있다.The food composition may be a health food composition, and the formulation of the food composition is not particularly limited, but may be formulated as, for example, tablets, pills, soft and hard capsules, granules, drinks, caramels, powders, and the like. In the food composition of each formulation, in addition to the active ingredients, ingredients commonly used in the field may be appropriately selected and blended by a person skilled in the art according to the formulation or purpose of use without difficulty, and synergistic effects may occur when applied simultaneously with other ingredients. The determination of the dosage of the active ingredient is within the level of those skilled in the art, and the daily dosage thereof is, for example, 10 5 to 10 13 CFU/day, more specifically 10 8 to 10 10 CFU/day for Lactobacillus genseni. It may be, but is not limited thereto, and may vary according to various factors such as age, health condition, and complications of the target to be administered.
항균용 미생물 제제의 제조 방법Method for producing antibacterial microbial preparation
본 발명은 락토바실러스 젠세니 SWPM104 균주를 배양하는 단계를 포함하는 항균용 미생물 제제의 제조 방법에 대한 것이다.The present invention relates to a method for producing an antimicrobial microbial preparation comprising culturing the Lactobacillus genseni SWPM104 strain.
화장료 조성물Cosmetic composition
본 발명은 락토바실러스 젠세니 SWPM104, 그 배양물 또는 그 추출물을 포함하는, 화장료 조성물에 대한 것이다. 상기 화장료 조성물은 모든 제형으로 제공될 수 있다. 예를 들면, 액상, 에멀젼, 현탁액, 고체, 겔, 분말, 페이스트, 폼 에어로졸 또는 패치 조성물의 제형으로 제공될 수 있다. 이러한 제형의 조성물은 당해 분야의 통상적인 방법에 따라 제조될 수 있다.The present invention relates to a cosmetic composition comprising Lactobacillus genseni SWPM104, its culture or its extract. The cosmetic composition may be provided in any formulation. For example, it may be provided in the form of a liquid, emulsion, suspension, solid, gel, powder, paste, foam aerosol or patch composition. Compositions of such formulations can be prepared according to conventional methods in the art.
상기 화장료 조성물은 상기한 물질 이외에 주 효과를 손상시키지 않는 범위 내에서, 바람직하게는 주 효과에 상승 효과를 줄 수 있는 다른 성분들을 함유할 수 있다. 본 발명에 따른 화장료 조성물은 비타민, 아미노산, 고분자 펩타이드, 지질로 이루어진 군에서 선택된 물질을 포함할 수 있다. 또한 본 발명에 따른 화장료 조성물은 보습제, 에몰리언트제, 계면 활성제, 자외선 흡수제, 방부제, 살균제, 산화 방지제, pH 조정제, 유기 및 무기 안료, 향료를 포함할 수 있다. 상기 성분의 배합량은 본 발명의 목적 및 효과를 손상시키지 않는 범위 내에서 당업자가 용이하게 선정 가능하며, 그 배합량은 조성물 전체 중량을 기준으로 0.01~5 중량%, 구체적으로 0.01~3 중량%일 수 있다.In addition to the above-described substances, the cosmetic composition may contain other ingredients that can give a synergistic effect to the main effect within a range that does not impair the main effect. The cosmetic composition according to the present invention may include a substance selected from the group consisting of vitamins, amino acids, polymer peptides, and lipids. In addition, the cosmetic composition according to the present invention may include a moisturizing agent, an emollient agent, a surfactant, an ultraviolet absorber, a preservative, a bactericide, an antioxidant, a pH adjuster, an organic and inorganic pigment, and a fragrance. The blending amount of the ingredients can be easily selected by a person skilled in the art within a range that does not impair the object and effect of the present invention, and the blending amount can be 0.01 to 5% by weight, specifically 0.01 to 3% by weight, based on the total weight of the composition. have.
사료 조성물Feed composition
본 발명은 락토바실러스 젠세니 SWPM104, 그 배양물 또는 그 추출물을 포함하는, 사료 조성물에 대한 것이다.The present invention relates to a feed composition comprising Lactobacillus genseni SWPM104, its culture or its extract.
본 발명의 조성물The composition of the present invention
본 발명은 락토바실러스 젠세니 SWPM104, 그 배양물 또는 그 추출물을 포함하는 조성물에 대한 것이다. 이 때, 상기 배양물 또는 추출물은 락토바실러스 젠세니 SWPM104 균주의 생균 또는 사균, 상기 균주의 파쇄물, 상기 균주의 배양물, 상기 배양물의 농축액, 상기 배양물의 건조물 및 상기 배양물의 추출물 등을 포함한다. 상기 사균은 본 발명의 락토바실러스 젠세니(Lactobacillus jensenii) SWPM104을 배양한 후 가열, 냉각 등의 온도, 초음파, 효소, 압력, 산 및 염기 등의 처리를 통해 생균을 사멸시킨 사균으로서, 균주의 발효에 의한 대사산물을 포함하는 배양액을 포함하거나 포함하지 않을 수 있다. 발효에 의한 배양액을 포함시키지 않을 경우 발효시킨 배양액을 여과 또는 원심 분리하여 균체를 회수한 후 생균을 사멸시키거나, 먼저 사멸시킨 후 배양액으로부터 사멸된 균체를 회수할 수도 있다. 본 발명의 균주를 배양하는 방법은 당업계에 통상적으로 이용되는 방법에 따라 배양할 수 있다. 상기 균주 배양물의 추출물은 균주 배양물의 용매 추출물이 수 있다. 상기 용매는 당업계에서 일반적으로 사용하는 용매이면 되고, 특별히 제한되지 않는다. 예컨대, 상기 용매는 에틸 아세테이트일 수 있다. 상기 배양물은 락토바실러스 젠세니 SWPM104 균주를 배양 배지 또는 배양액에서 배양한 것을 의미하며, 상기 배양물은 본 발명의 균주를 포함하거나 포함하지 않을 수 있다. 상기 배양물은 그 제형이 액체 또는 고체일 수 있으나, 이에 제한되는 것은 아니다.The present invention relates to a composition comprising Lactobacillus genseni SWPM104, a culture thereof or an extract thereof. In this case, the culture or extract includes live or dead bacteria of Lactobacillus genseni SWPM104 strain, a lysate of the strain, a culture of the strain, a concentrate of the culture, a dried product of the culture, and an extract of the culture. The dead cells are the dead cells obtained by culturing the Lactobacillus jensenii SWPM104 of the present invention and then killing live cells through treatment of temperature such as heating and cooling, ultrasonication, enzymes, pressure, acids and bases, and fermentation of the strain. It may or may not contain a culture medium containing metabolites by. If the fermented culture medium is not included, the fermented culture medium may be filtered or centrifuged to recover the cells, and then live cells may be killed, or the dead cells may be recovered from the culture solution after first killing. The method of culturing the strain of the present invention can be cultured according to a method commonly used in the art. The extract of the strain culture may be a solvent extract of the strain culture. The solvent may be a solvent generally used in the art, and is not particularly limited. For example, the solvent may be ethyl acetate. The culture means that the Lactobacillus genseni SWPM104 strain is cultured in a culture medium or a culture medium, and the culture may or may not contain the strain of the present invention. The culture may have a liquid or solid formulation, but is not limited thereto.
본 발명의 이점 및 특징, 그리고 그것들을 달성하는 방법은 상세하게 후술되어 있는 실시예들을 참조하면 명확해질 것이다. 그러나, 본 발명은 이하에서 개시되는 실시예들에 한정되는 것이 아니라 서로 다른 다양한 형태로 구현될 것이며, 단지 본 실시예들은 본 발명의 개시가 완전하도록 하며, 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자에게 발명의 범주를 완전하게 알려주기 위해 제공되는 것이며, 본 발명은 청구항의 범주에 의해 정의될 뿐이다.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, only these embodiments make the disclosure of the present invention complete, and common knowledge in the technical field to which the present invention pertains. It is provided to completely inform the scope of the invention to those who have, and the invention is only defined by the scope of the claims.
<재료 및 방법><Materials and methods>
비교 균주로는 본 발명자가 보유하고 있던 락토바실러스 살리바리우스 SWPM101 (KCTC13436BP) 및 락토바실러스 가세리 SWPM102 (KCTC13437BP)를 사용하였다.As a comparative strain, Lactobacillus salivarius SWPM101 (KCTC13436BP) and Lactobacillus gaseri SWPM102 (KCTC13437BP) owned by the present inventor were used.
<실험예 1> 락토바실러스 젠세니 SWPM104 균주의 분리 및 동정<Experimental Example 1> Isolation and identification of Lactobacillus genseni SWPM104 strain
건양대병원 산부인과에 정기검진을 받기 위해 내원한 건강한 한국인 임산부들의 동의를 받아 이들 임산부들의 질내 시료를 채취하였다. 상기 시료로부터 수득한 균주를 1.5% 한천(agar)이 포함된 MRS 고체 배지 (Difco, USA)에 도말하여 37℃에서 48시간, GasPak (BD, USA)을 이용해 혐기 조건에서 배양하였다. 그리고 순수 분리된 집락을 백금이(loop)로 취하여 MRS 액체 배지(Difco, USA)에서 37 ℃, 48시간, 혐기배양 하였다.With the consent of healthy Korean pregnant women who visited Konyang University Hospital for regular checkups, vaginal samples were collected from these pregnant women. The strain obtained from the sample was plated on MRS solid medium (Difco, USA) containing 1.5% agar, and cultured at 37° C. for 48 hours using GasPak (BD, USA) under anaerobic conditions. Then, the purely separated colonies were taken with a loop and incubated anaerobic at 37° C. for 48 hours in MRS liquid medium (Difco, USA).
그런 다음, 배양된 균주의 동정 및 분류를 위하여 16s rRNA 유전자 염기서열 및 Miseq을 이용하여 초안 유전체 정보를 분석하였다. 상기 균주의 16s rRNA 유전자의 염기서열 결정(서열번호 1) 및 분석 결과는 표 1과 같다. 16s rRNA 염기서열 분석 결과, 상기 균주는 락토바실러스 젠세니 (Lactobacillus jensenii strain Gasser 62G, DSM 20557, ATCC 25258)와 가장 높은 상동성(99%)을 나타내어 락토바실러스 젠세니 (Lactobacillus jensenii)로 동정하였다. 상기 균주를 락토바실러스 젠세니 SWPM104로 명명하였으며, 생명공학연구원 유전자은행에 2020년 4월 8일자로 기탁하였다 (수탁번호 KCTC14168BP).Then, for identification and classification of the cultured strains, the draft genome information was analyzed using the 16s rRNA gene sequence and Miseq. Table 1 shows the base sequence determination (SEQ ID NO: 1) and analysis results of the 16s rRNA gene of the strain. As a result of 16s rRNA sequencing, the strain showed the highest homology (99%) with Lactobacillus jensenii strain Gasser 62G, DSM 20557, ATCC 25258, and was identified as Lactobacillus jensenii . The strain was named Lactobacillus genseni SWPM104, and was deposited with the Institute of Biotechnology Gene Bank on April 8, 2020 (accession number KCTC14168BP).
<실험예 2> 내산성 평가<Experimental Example 2> Acid resistance evaluation
락토바실러스 젠세니 SWPM104 균주의 내산성을 평가하였다. 평가 방법은 하기와 같다. 균주들을 MRS 액체 배지(pH 7)를 pH 2와 pH 3으로 적정하여 만든 산성 액체 배지에서 10-6 배율로 희석하였다. 희석한 균액을 100 μL씩 도말하여 24시간 동안 37℃에서 혐기 배양한 뒤 콜로니 수를 측정하면서 시간대별로 MRS agar 배지에 도말하여 48시간 동안 혐기적으로 배양한 후 콜로니 수를 측정하였다. Acid resistance of the Lactobacillus genseni SWPM104 strain was evaluated. The evaluation method is as follows. The strains were diluted at 10 -6 magnification in an acidic liquid medium prepared by titrating MRS liquid medium (pH 7) to
그 결과, 락토바실러스 젠세니 SWPM104 균주는 pH 3의 배양 조건에서 성장이 우수한 것으로 나타났다. 그러므로 락토바실러스 젠세니 SWPM104 균주는 내산성을 가지고 있는 것이 확인되었다(표 2).As a result, the Lactobacillus genseni SWPM104 strain was found to be excellent in growth in the culture conditions of
<실험예 3> 내담즙성 평가<Experimental Example 3> Evaluation of bile resistance
락토바실러스 젠세니 SWPM104 균주의 내담즙성을 평가하였다. 이때 비교 균주로는 락토바실러스 젠세니 GASSER 62G 균주를 사용하였다. 평가 방법은 하기와 같다. 0.3 중량%, 1 중량 %의 담즙산염이 포함된 배지에 균주를 37℃, 24시간 혐기 배양하면서 MRS agar 배지에 도말하여 48시간 동안 혐기 배양한 후 콜로니 수를 측정하였다. Bile resistance of the Lactobacillus genseni SWPM104 strain was evaluated. At this time, Lactobacillus genseni GASSER 62G strain was used as a comparative strain. The evaluation method is as follows. The strain was plated on MRS agar medium while anaerobic culture at 37° C. for 24 hours in a medium containing 0.3% by weight and 1% by weight of bile salt, followed by anaerobic culture for 48 hours, and then the number of colonies was measured.
그 결과, 락토바실러스 젠세니 SWPM104 균주는 0.3 중량% 담즙산염 농도에서, 우수한 성장력을 보였으며, 1 중량%의 담즙산염 농도에서는 모두 사멸하였다. 0.3 중량% 농도의 담즙농도는 사람 소장 내 평균 담즙 농도이므로 락토바실러스 젠세니 SWPM104 균주를 경구 섭취하는 경우에도 상기 균주는 높은 생존율을 유지할 것으로 판단되었다 (표 3, 도 1).As a result, the Lactobacillus genseni SWPM104 strain showed excellent growth at a concentration of 0.3% by weight bile salt, and all died at a concentration of 1% by weight bile salt. Since the bile concentration of 0.3% by weight is the average bile concentration in the human small intestine, even when the Lactobacillus genseni SWPM104 strain was taken orally, the strain was determined to maintain a high survival rate (Table 3, FIG. 1).
<실험예 4> 항생제 감수성 평가<Experimental Example 4> Antibiotic sensitivity evaluation
기능성 식품 소재 등으로 활용시의 안전성을 확인하기 위하여 락토바실러스 젠세니 SWPM104 균주에 대하여 항생제 저항성을 평가하였다. 현재 락토바실러스 계열 유산균의 식품 활용시의 항생제 저항성에 관한 규격기준은 설정되어 있지 않는 바, 동물성 사료에 첨가되는 미생물의 항생제 저항성에 관한 국제 기준인 EFSA 기준을 바탕으로 평가하였다.Antibiotic resistance was evaluated against Lactobacillus genseni SWPM104 strain to confirm the safety when used as a functional food material. Currently, the standard for antibiotic resistance of Lactobacillus family lactic acid bacteria in food use is not established, so the evaluation was based on the EFSA standard, an international standard for antibiotic resistance of microorganisms added to animal feed.
구체적으로, 락토바실러스 젠세니 SWPM104 균주의 항생제 감수성 평가는 국제 기준인 EFSA (European Food Safety Authority) 지침에 의거하여, ampicillin (AMP), chloramphenicol (CHR), clindamycin(CLM), erythromycin (ERY), gentamycin (GEN), kanamycin(KAN), streptomycin (STR), tetracylcine (TET), vancomycin(VAN) 등 9종의 항생제를 대상으로 실시하였다. 항생제 저항성 평가 시에 사용된 시험방법은 유산균의 항생제 저항성 시험을 위한 SOP 기준으로 실시하였고, LSM-Broth에 ~6 × 106 CFU/ml로 락토바실러스 젠세니 SWPM104 균주를 접종한 후, 각 항생제 별 MIC test strip을 올려놓고 37℃에서 48시간 동안 혐기배양 후 억제 정도 및 MIC를 평가하였다.Specifically, the evaluation of antibiotic susceptibility of Lactobacillus genseni SWPM104 strain is based on the international standard EFSA (European Food Safety Authority) guidelines, ampicillin (AMP), chloramphenicol (CHR), clindamycin (CLM), erythromycin (ERY), gentamycin (GEN), kanamycin (KAN), streptomycin (STR), tetracylcine (TET), and vancomycin (VAN) were tested for 9 antibiotics. The test method used in the evaluation of antibiotic resistance was conducted as the SOP standard for the antibiotic resistance test of lactic acid bacteria, and after inoculating LSM-Broth with Lactobacillus genseni SWPM104 strain at ~6 × 10 6 CFU/ml, each antibiotic The MIC test strip was placed and the degree of inhibition and MIC were evaluated after anaerobic culture at 37°C for 48 hours.
그 결과, 락토바실러스 젠세니 SWPM104 균주는 사용된 9종의 항생제에 대하여 EFSA의 기준(표 4의 EFSA guideline)을 만족하는 항생제 감수성을 나타내었으며, 경구 섭취를 통한 건강 보조 식품으로서의 활용 가능성이 기대되었다(표 4, 도 2).As a result, the Lactobacillus genseni SWPM104 strain showed antibiotic susceptibility that satisfies the EFSA criteria (EFSA guideline in Table 4) with respect to the nine antibiotics used, and is expected to be used as a health supplement through oral intake. (Table 4, Fig. 2).
<실험예 5> 과산화수소 생성능 평가<Experimental Example 5> Evaluation of hydrogen peroxide production ability
고체 배양된 균주 plate를 30분간 산소에 노출시킨 후 colony 색이 푸른색으로 변색이 되는 정도에 따라 다음과 같이 분류하였다다. 육안으로 관찰하여 30분 이내에 colony가 변색이 되지 않으면 (-), 일부 colony만 변색이 되면 1급 (+), 전체적으로 변색 되었으나 색이 옅으면 2급 (++), colony가 진한 푸른색을 나타내면 3급 (+++)으로 등급을 분류하였다.After exposing the solid cultured strain plate to oxygen for 30 minutes, the colony color was classified as follows according to the degree of discoloration to blue. If the colony does not discolor within 30 minutes by visual observation, it is (-), if only some colonies are discolored, grade 1 (+), if the color is completely discolored, grade 2 (++), and if the colony is dark blue Classified as 3 grade (+++).
그 결과, 락토바실러스 젠세니 SWPM104 균주는 2급(++)으로 나타났으며, 과산화수소 생성력을 갖는 것을 확인하였다. (도 3)As a result, it was confirmed that the Lactobacillus genseni SWPM104 strain was grade 2 (++), and had hydrogen peroxide producing power. (Fig. 3)
<실험예 6> 항균력 평가<Experimental Example 6> Evaluation of antibacterial activity
가르드네렐라 바기날리스(Gardnerella vaginalis), 아토포비움 바기날리스(Atopobium vaginalis), 스트렙토쿠스 뮤탄스(Streptococcus mutans), 녹농균(Pseudomonas aeruginosa), 표피포도구균(Staphylococcus epidermidis), 바실러스 세레우스(Bacillus cereus) 및 살모넬라 엔테리티디스(Salmonella enteritidis)에 대한 락토바실러스 젠세니 SWPM104 균주의 항균력을 평가하였다.Gardnerella vaginalis, Atopobium vaginalis, Streptococcus mutans, Pseudomonas aeruginosa, Staphylocilloccus epidermidis, Bacillus cereus cereus) and Salmonella enteritidis (Salmonella enteritidis) were evaluated for the antibacterial activity of Lactobacillus genseni SWPM104 strain.
이때, 락토바실러스 젠세니 SWPM104 균주의 배양액의 항균력을 측정하기 위하여 KS 항균력 시험방법을 사용하여 평가하였다. 평가 방법은 하기와 같다. 가르드네렐라 바기날리스(Gardnerella vaginalis), 아토포비움 바기날리스(Atopobium vaginalis), 스트렙토쿠스 뮤탄스(Streptococcus mutans), 녹농균(Pseudomonas aeruginosa), 표피포도구균(Staphylococcus epidermidis), 바실러스 세레우스(Bacillus cereus) 및 살모넬라 엔테리티디스(Salmonella enteritidis), 대장균(Escherichia coli) 배양액을 각각 1.0mL씩 채취하여 250mL 용량의 멸균 삼각 플라스크에 접종하여 (37±1) ℃에서 24시간 진탕 배양하였다. 배양 용액의 균수를 흡광 강도계로 측정하여 추정한 후 희석 용액은 TSB배지에서 균 수가 1-5 × 105 개 (CFU/mL)가 되도록 희석 조제하여 사용하였다. 시료를 가로 세로 각각 1.0cm × 1.0cm 크기로 준비하고 70% 에탄올로 살균하여 (105±1)℃의 건조기에서 30분간 건조하거나 자외선 (UV254nm)살균기에서 2시간 이상 살균하여 사용하였다. 한편, 시험 시료와 동일한 크기 또는 양으로 균들을 채취하여 항균 가공 시험 시료와 동일하게 처리한 검체를 준비하고, 이것을 대조 시료로 사용하였다. 미리 조정하여 둔 각각의 시험 용액에 시험 시료와 대조 시료를 넣고 균 배양액을 1.0mL씩 접종하여 균일하게 혼합하고 접종 즉시 시험 시료와 대조 시료를 포함하고 있는 삼각 플라스크로부터 균 약을 일정량 채취하여 생리 식염수로 희석하고, TSA 배지를 사용하여 (37±1)℃에서 24시간 배양 후 균수를 확인하였다. 이때 희석 비율은 101 내지 104의 희석 범위로 하였다. 시험 시료에 의한 세균의 감소율은 하기 식 1에 따라 계산하였다.At this time, in order to measure the antibacterial activity of the culture medium of Lactobacillus genseni SWPM104 strain, it was evaluated using the KS antibacterial activity test method. The evaluation method is as follows. Gardnerella vaginalis, Atopobium vaginalis, Streptococcus mutans, Pseudomonas aeruginosa, Staphylocilloccus epidermidis, Bacillus cereus cereus), Salmonella enteritidis, and Escherichia coli cultures were collected at 1.0 mL each, inoculated into a 250 mL sterile Erlenmeyer flask, and cultured with shaking at (37±1)° C. for 24 hours. After estimating the number of bacteria in the culture solution by measuring it with an absorption intensity meter, the diluted solution was prepared and diluted so that the number of bacteria was 1-5 × 10 5 (CFU/mL) in TSB medium. Samples were prepared in dimensions of 1.0cm × 1.0cm in width and length, respectively, and sterilized with 70% ethanol, dried for 30 minutes in a dryer at (105±1)°C, or sterilized in an ultraviolet (UV254nm) sterilizer for 2 hours or longer. On the other hand, a sample treated in the same manner as the antibacterial processing test sample was prepared by collecting bacteria in the same size or amount as the test sample, and this was used as a control sample. Add the test sample and the control sample to each pre-adjusted test solution, inoculate 1.0 mL of the fungal culture solution, and mix uniformly. Immediately after inoculation, a certain amount of the fungal drug is collected from the Erlenmeyer flask containing the test sample and the control sample and physiological saline solution. Diluted with, and cultured for 24 hours at (37±1)°C using TSA medium, the number of bacteria was confirmed. At this time, the dilution ratio was in the range of 10 1 to 10 4 . The reduction rate of bacteria by the test sample was calculated according to
<식 1><
이때,At this time,
Mb: 24시간 배양 후 대조 시료의 균수 (평균값)Mb: number of bacteria in the control sample after 24 hours incubation (average value)
Mc: 24시간 배양 후 시험 시료의 균수 (평균값)Mc: Number of bacteria in the test sample after incubation for 24 hours (average value)
그 결과, 락토바실러스 젠세니 SWPM104 균주는 질염의 원인균인 가르드네렐라 바기날리스 및 아토포비움 바기날리스, 충치의 원인균인 스트렙토쿠스 뮤탄스, 아토피성 피부염의 원인균인 표피포도구균 그리고 녹농균, 식중독균, 장염에 대하여 항균력이 있는 것이 확인되었다. 그러나 대장균에 대하여는 항균력이 없는 것으로 나타났다(표 5).As a result, Lactobacillus genseni SWPM104 strains were Gardnerella baginalis and Atofobium baginalis, which are the causative agents of vaginitis, Streptoccus mutans, which is the causative agent of tooth decay, Epidermal aureus and Pseudomonas aeruginosa, food poisoning bacteria , It was confirmed that it has antibacterial activity against enteritis. However, it was found that there was no antibacterial activity against E. coli (Table 5).
기탁기관명 : 한국생명공학연구원Name of donated institution: Korea Research Institute of Bioscience and Biotechnology
수탁번호 : KCTC14168BPAccession number: KCTC14168BP
수탁일자 : 20200408Consignment Date: 20200408
<110> Sungwun Pharmacopia Corporation <120> LACTOBACILLUS JENSENII SWPM104 WHICH HAS ANTIBACTERIAL ACTIVITY <130> KNP200009 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1431 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence of 16s rRNA of Lactobacillus jensenii <400> 1 agtcgagcga gcttgcctat agaaattctt cggaatggac atagatacaa gctagcggcg 60 gatgggtgag taacgcgtgg gtaacctgcc cttaagtctg ggataccatt tggaaacaga 120 tgctaatacc ggataaaagc tactttcgca tgaaagaagt ttaaaaggcg gcgtaagctg 180 tcgctaaagg atggacctgc gatgcattag ctagttggta aggtaacggc ttaccaaggc 240 gatgatgcat agccgagttg agagactgat cggccacatt gggactgaga cacggcccaa 300 actcctacgg gaggcagcag tagggaatct tccacaatgg acgaaagtct gatggagcaa 360 cgccgcgtga gtgaagaagg ttttcggatc gtaaagctct gttgttggtg aagaaggata 420 gaggtagtaa ctggccttta tttgacggta atcaaccaga aagtcacggc taactacgtg 480 ccagcagccg cggtaatacg taggtggcaa gcgttgtccg gatttattgg gcgtaaagcg 540 agcgcaggcg gattgataag tctgatgtga aagccttcgg ctcaaccgaa gaactgcatc 600 agaaactgtc aatcttgagt gcagaagagg agagtggaac tccatgtgta gcggtggaat 660 gcgtagatat atggaagaac accagtggcg aaggcggctc tctggtctgt aactgacgct 720 gaggctcgaa agcatgggta gcgaacagga ttagataccc tggtagtcca tgccgtaaac 780 gatgagtgct aagtgttggg aggtttccgc ctctcagtgc tgcagctaac gcattaagca 840 ctccgcctgg ggagtacgac cgcaaggttg aaactcaaag gaattgacgg gggcccgcac 900 aagcggtgga gcatgtggtt taattcgaag caacgcgaag aaccttacca ggtcttgaca 960 tcctttgacc acctaagaga ttaggttttc ccttcgggga caaagagaca ggtggtgcat 1020 ggctgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag cgcaaccctt 1080 gttaatagtt gccagcatta agttgggcac tctattgaga ctgccggtga caaaccggag 1140 gaaggtgggg atgacgtcaa gtcatcatgc cccttatgac ctgggctaca cacgtgctac 1200 aatgggcagt acaacgagaa gcgaacctgt gaaggcaagc ggatctctta aagctgttct 1260 cagttcggac tgtaggctgc aactcgccta cacgaagctg gaatcgctag taatcgcgga 1320 tcagcacgcc gcggtgaata cgttcccggg ccttgtacac accgcccgtc acaccatgag 1380 agtttgtaac acccaaagtc ggtgaggtaa ccttggagcc agccgcctaa g 1431 <110> Sungwun Pharmacopia Corporation <120> LACTOBACILLUS JENSENII SWPM104 WHICH HAS ANTIBACTERIAL ACTIVITY <130> KNP200009 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1431 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence of 16s rRNA of Lactobacillus jensenii <400> 1 agtcgagcga gcttgcctat agaaattctt cggaatggac atagatacaa gctagcggcg 60 gatgggtgag taacgcgtgg gtaacctgcc cttaagtctg ggataccatt tggaaacaga 120 tgctaatacc ggataaaagc tactttcgca tgaaagaagt ttaaaaggcg gcgtaagctg 180 tcgctaaagg atggacctgc gatgcattag ctagttggta aggtaacggc ttaccaaggc 240 gatgatgcat agccgagttg agagactgat cggccacatt gggactgaga cacggcccaa 300 actcctacgg gaggcagcag tagggaatct tccacaatgg acgaaagtct gatggagcaa 360 cgccgcgtga gtgaagaagg ttttcggatc gtaaagctct gttgttggtg aagaaggata 420 gaggtagtaa ctggccttta tttgacggta atcaaccaga aagtcacggc taactacgtg 480 ccagcagccg cggtaatacg taggtggcaa gcgttgtccg gatttattgg gcgtaaagcg 540 agcgcaggcg gattgataag tctgatgtga aagccttcgg ctcaaccgaa gaactgcatc 600 agaaactgtc aatcttgagt gcagaagagg agagtggaac tccatgtgta gcggtggaat 660 gcgtagatat atggaagaac accagtggcg aaggcggctc tctggtctgt aactgacgct 720 gaggctcgaa agcatgggta gcgaacagga ttagataccc tggtagtcca tgccgtaaac 780 gatgagtgct aagtgttggg aggtttccgc ctctcagtgc tgcagctaac gcattaagca 840 ctccgcctgg ggagtacgac cgcaaggttg aaactcaaag gaattgacgg gggcccgcac 900 aagcggtgga gcatgtggtt taattcgaag caacgcgaag aaccttacca ggtcttgaca 960 tcctttgacc acctaagaga ttaggttttc ccttcgggga caaagagaca ggtggtgcat 1020 ggctgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag cgcaaccctt 1080 gttaatagtt gccagcatta agttgggcac tctattgaga ctgccggtga caaaccggag 1140 gaaggtgggg atgacgtcaa gtcatcatgc cccttatgac ctgggctaca cacgtgctac 1200 aatgggcagt acaacgagaa gcgaacctgt gaaggcaagc ggatctctta aagctgttct 1260 cagttcggac tgtaggctgc aactcgccta cacgaagctg gaatcgctag taatcgcgga 1320 tcagcacgcc gcggtgaata cgttcccggg ccttgtacac accgcccgtc acaccatgag 1380 agtttgtaac acccaaagtc ggtgaggtaa ccttggagcc agccgcctaa g 1431
Claims (12)
Lactobacillus jensenii having antibacterial activity against Streptococcus mutans SWPM104 (Accession No.: KCTC 14168BP).
상기 락토바실러스 젠세니 SWPM104은 가르드네렐라 바기날리스(Gardnerella vaginalis), 아토포비움 바기날리스(Atopobium vaginalis), 녹농균(Pseudomonas aeruginosa), 표피포도구균(Staphylococcus epidermidis), 바실러스 세레우스(Bacillus cereus) 및 살모넬라 엔테리티디스(Salmonella enteritidis)으로 구성되는 군으로부터 선택되는 하나 이상에 대하여 항균 활성 갖는 것을 특징으로 하는 락토바실러스 젠세니 SWPM104.
The method of claim 1,
The Lactobacillus genseni SWPM104 is Gardnerella vaginalis, Atopobium vaginalis, Pseudomonas aeruginosa, Staphylococcus epidermidis, Bacillus cereus And Salmonella enteritidis (Salmonella enteritidis) Lactobacillus genseni SWPM104, characterized in that it has antibacterial activity against at least one selected from the group consisting of.
상기 락토바실러스 젠세니 SWPM104은 대장균(Escherichia coli)에 대하여 항균 활성이 없는 것을 특징으로 하는 락토바실러스 젠세니 SWPM104.
The method of claim 1,
The Lactobacillus genseni SWPM104 is Lactobacillus genseni SWPM104, characterized in that it has no antibacterial activity against Escherichia coli.
상기 락토바실러스 젠세니 SWPM104은 pH3의 조건 하 증식능을 갖는 것을 특징으로 하는 락토바실러스 젠세니 SWPM104.
The method of claim 1,
The Lactobacillus genseni SWPM104 is Lactobacillus genseni SWPM104, characterized in that it has a proliferative ability under conditions of pH3.
상기 락토바실러스 젠세니 SWPM104은 pH7의 조건 하 증식능을 갖는 것을 특징으로 하는 락토바실러스 젠세니 SWPM104.
The method of claim 1,
The Lactobacillus genseni SWPM104 is Lactobacillus genseni SWPM104, characterized in that it has a proliferative ability under conditions of pH7.
상기 락토바실러스 젠세니 SWPM104은 0.3 중량% 담즙산을 포함하는 배지 조건 하 증식능을 갖는 것을 특징으로 하는 락토바실러스 젠세니 SWPM104.
The method of claim 1,
The Lactobacillus genseni SWPM104 is Lactobacillus genseni SWPM104, characterized in that it has a proliferative ability under a medium condition containing 0.3% by weight bile acid.
A food composition comprising the Lactobacillus genseni SWPM104 of claim 1, a culture thereof or an extract thereof.
상기 식품 조성물은 가르드네렐라 바기날리스(Gardnerella vaginalis), 아토포비움 바기날리스(Atopobium vaginalis), 스트렙토쿠스 뮤탄스(Streptococcus mutans), 녹농균(Pseudomonas aeruginosa), 표피포도구균(Staphylococcus epidermidis), 바실러스 세레우스(Bacillus cereus) 및 살모넬라 엔테리티디스(Salmonella enteritidis)으로 구성되는 군으로부터 선택되는 하나 이상의 세균에 대하여 항균 활성을 갖는 것을 특징으로 하는 식품 조성물.
The method of claim 8,
The food composition is Gardnerella vaginalis, Atopobium vaginalis, Streptococcus mutans, Pseudomonas aeruginosa, Staphylococcus epidermidis, and Bacillus. Cereus (Bacillus cereus) and Salmonella enteritidis (Salmonella enteritidis) food composition characterized in that it has an antibacterial activity against one or more bacteria selected from the group consisting of.
Claim 1 Lactobacillus genseni SWPM104, including the culture or its extract, Gardnerella vaginalis, Atopobium vaginalis, Streptococcus mutans, Pharmaceutical for the prevention or treatment of diseases caused by one or more bacteria selected from the group consisting of Pseudomonas aeruginosa, Staphylococcus epidermidis, Bacillus cereus, and Salmonella enteritidis. As a composition, the disease is at least one disease selected from the group consisting of bacterial vaginosis, atopic dermatitis, respiratory disease due to Pseudomonas aeruginosa infection, pneumonia and tooth decay due to Pseudomonas aeruginosa infection, pharmaceutical composition.
Claim 1 Lactobacillus genseni SWPM104, comprising a culture or an extract thereof, feed composition.
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| CN116042432A (en) * | 2022-07-14 | 2023-05-02 | 沈阳医学院 | Novel strain with broad-spectrum antibacterial property and purification of antibacterial substances thereof |
| CN116376758A (en) * | 2023-03-15 | 2023-07-04 | 广东南芯医疗科技有限公司 | Lactobacillus jensenii LS06 and application thereof in preparation of medicines for treating or preventing allergic diseases |
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