KR20040044300A - Composition containing Lactococcus lactis NK24 (KFCC 11315) for stomach disorder - Google Patents

Composition containing Lactococcus lactis NK24 (KFCC 11315) for stomach disorder Download PDF

Info

Publication number
KR20040044300A
KR20040044300A KR1020020072758A KR20020072758A KR20040044300A KR 20040044300 A KR20040044300 A KR 20040044300A KR 1020020072758 A KR1020020072758 A KR 1020020072758A KR 20020072758 A KR20020072758 A KR 20020072758A KR 20040044300 A KR20040044300 A KR 20040044300A
Authority
KR
South Korea
Prior art keywords
kfcc
lactis
bacteria
pylori
lactococcus lactis
Prior art date
Application number
KR1020020072758A
Other languages
Korean (ko)
Other versions
KR100518263B1 (en
Inventor
백현동
이나경
최신양
Original Assignee
학교법인 한마학원
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 학교법인 한마학원 filed Critical 학교법인 한마학원
Priority to KR10-2002-0072758A priority Critical patent/KR100518263B1/en
Publication of KR20040044300A publication Critical patent/KR20040044300A/en
Application granted granted Critical
Publication of KR100518263B1 publication Critical patent/KR100518263B1/en

Links

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66Microorganisms or materials therefrom
    • A61K35/74Bacteria
    • A61K35/741Probiotics
    • A61K35/744Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Mycology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Microbiology (AREA)
  • Epidemiology (AREA)
  • Molecular Biology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

PURPOSE: Provided is a composition containing Lactococcus lactis NK24 (KFCC 11315) having excellent antibacterial effect, as an active ingredient, for the prevention and treatment of stomach disorder. CONSTITUTION: A composition for the prevention and treatment of stomach disorder is characterized by containing Lactococcus lactis NK24 (KFCC 11315), as an active ingredient. It has excellent antibacterial effect on helicobacter pylori and tolerance to gastric and bile juices, and produce beta-galactosidase to reduce lactose intolerance.

Description

락토코쿠스 락티스 NK24 (KFCC 11315) 생균을 유효성분으로 함유하는 것을 특징으로 하는 위장병 예방 및 치료용 조성물 {Composition containing Lactococcus lactis NK24 (KFCC 11315) for stomach disorder}[Composition containing Lactococcus lactis NK24 (KFCC 11315) for stomach disorder} comprising a probiotic Lactococcus lactis NX24 (JPFC 11315) as an active ingredient

본 발명은 헬리코박터 파이로리(H. pylori) 균주에 대해서 항균효과가 우수한 생균을 유효성분으로 함유하는 것을 특징으로 하는 위장병 예방 및 치료용 조성물에 관한 것으로, 더욱 상세하게는 헬리코박터 파이로리(H. pylori) 균주에 대해서 우수한 항균효과가 있는 락토코쿠스 락티스(Lactococcus lactis) NK24 (KFCC 11315) 생균을 유효성분으로 함유하는 것을 특징으로 하는 위장병 예방 및 치료용 조성물에 관한 것이다.The invention Helicobacter pylori (H. pylori) strains for the present invention relates to the prevention and treatment of gastrointestinal composition characterized by containing as an active ingredient the antibacterial effect excellent live cells, and more particularly, to Helicobacter pylori (H. pylori) strains It relates to a composition for the prevention and treatment of gastrointestinal diseases, which contains Lactococcus lactis NK24 (KFCC 11315) live bacteria having an excellent antimicrobial effect as an active ingredient.

소득 수준이 향상되면서 소비자는 건강 지향적인 제품에 관심을 가지고 있으며, 기능성 식품에 대한 수요 역시 크게 증가되고 있다. 실제로 프로바이오틱 생균의 기능성과 효능을 이용한 식품개발은 기능성 식품의 중요한 분야로 인식되고 있으며 기능성 식품으로 프로바이오틱 생균을 이용하려는 연구가 활발하게 이루어지고 있다. 프로바이오틱 생균제란 살아있는 미생물 균체를 섭취함으로써 미생물이 분비하는 효소, 유기산, 비타민 및 무독성 항균물질 등에 의한 장내 균총의 정상화는 물론 장질환 치료를 통해 신체기능 개선을 목적으로 생산된 제품을 말한다(Fuller, R. Probiotics in man and animals. J. Appl. Bacteriol. 66:365-37 (1989)). 이러한 프로바이오틱 생균제는 장내 균총의 안정화, 유해 세균의 정착 억제에 따른 부패산물 생성 감소 및 질병예방, 면역의 활성화, 간접적인 항암작용, 콜레스테롤 저하, 유당 불내증의 감소, 변비 억제 및 예방과 관련되어 기능성이 인정되어 있다(Goldin, B.R. and Borbach, S.L. Alterations in fecal microflora enzyme related to diet, age, lactobacillus supplements, and dimethylhydrazine. Cancer 40: 2421-2426 (1977); Gilliland, S.E. and Walker, D.K. Factors to consider when selecting a culture ofLactobacillus acidophilusas a dietary adjunct to produce a hypocholesterolemic effect in humans. J. Dairy Sci. 73: 905-911 (1990); Fuller, R. Probiotics in human medicine. Gut 32: 439-442 (1991); Hilton, E., Henry, D.I., Phyllis, A., Kenneth, F. and Michael, T.B. Ingestion of yoghurt containingLactobacillus acidophilusas prophylaxis for candidal vaginitis. Ann. Int. Med. 116: 353-357 (1992); Shah, N.Lactobacillus acidophilusand lactose intolerance, areview. ASEAN Food J. 9: 47-54 (1994); Matsuzaki, T., Hashimoto, S. and Yokokura, T. Effects on antitumor activity and cytokine production in the thoracic cavity by intrapleural administration ofLactobacillus caseiin tumor-bearing mice. Med. Microbiol. Immunol. Berl. 185: 157-161 (1996); Takiguchi, R., Mochizuki, E., Suzuki, Y., Nakajima, I. and Benno, Y.Lactobacillus acidophilusSBT 2062 andBifidobacterium longumSBT 2928 on harmful intestinal bacteria. J. Int. Microbiol. 11: 11-17 (1997)).As income levels rise, consumers are interested in health-oriented products, and the demand for functional foods is also increasing. In fact, the development of food using the functionality and efficacy of probiotic probiotics is recognized as an important field of functional foods, and researches on using probiotic probiotics as functional foods are being actively conducted. Probiotic probiotics are products that are produced for the purpose of improving the body function through the intestinal diseases as well as normalizing the intestinal flora by enzymes, organic acids, vitamins and non-toxic antibacterial substances secreted by microorganisms by ingesting live microbial cells , R. Probiotics in man and animals.J. Appl. Bacteriol. 66 : 365-37 (1989)). These probiotic probiotics are associated with stabilizing the intestinal flora, reducing rot production and preventing disease, preventing immunity, and preventing disease, activating immunity, indirect anticancer activity, lowering cholesterol, reducing lactose intolerance, and preventing and preventing constipation. Functionality is recognized (Goldin, BR and Borbach, SL Alterations in fecal microflora enzyme related to diet, age, lactobacillus supplements, and dimethylhydrazine. Cancer 40: 2421-2426 (1977); Gilliland, SE and Walker, DK Factors to consider when selecting a culture of Lactobacillus acidophilus as a dietary adjunct to produce a hypocholesterolemic effect in humans.J. Dairy Sci. 73: 905-911 (1990); Fuller, R. Probiotics in human medicine.Gut 32: 439-442 (1991 Hilton; E., Henry, DI, Phyllis, A., Kenneth, F. and Michael, TB Ingestion of yoghurt containing Lactobacillus acidophilus as prophylaxis for candidal vaginitis.Ann. Int. Med. 116: 353-357 (1992) Shah, N. Lactobacillus acidophil us and lactose intolerance, areview.ASEAN Food J. 9: 47-54 (1994); Matsuzaki, T., Hashimoto, S. and Yokokura, T. Effects on antitumor activity and cytokine production in the thoracic cavity by intrapleural administration of Lactobacillus casei in tumor-bearing mice. Med. Microbiol. Immunol. Berl. 185: 157-161 (1996); Takiguchi, R., Mochizuki, E., Suzuki, Y., Nakajima, I. and Benno, Y. Lactobacillus acidophilus SBT 2062 and Bifidobacterium longum SBT 2928 on harmful intestinal bacteria. J. Int. Microbiol. 11: 11-17 (1997).

상기와 같은 기능이 있는 프로바이오틱 생균제로 많이 이용되고 있는 세균은 락토바실러스(Lactobacillus)속,스트렙토코쿠스(Streptococcus)속, 비피도박테리움(Bifidobacterium)속, 엔테로코쿠스(Enterococcus)속, 클로스트리움 부티리쿰(Clostridium butyricum), 락토바실러스 스포로제네스(Lactobacillus sporogenes), 바실러스 서브틸리스(Bacillus subtilis) 바실러스 폴리퍼멘티쿠스(B. polyfermenticus) SCD 등이 있다.Bacteria that are widely used as probiotic probiotics with the above functions are Lactobacillus (Lactobacillus)genus,Streptococcus(StreptococcusGenus, Bifidobacterium (Bifidobacterium)genus, Enterococcus(EnterococcusClostridium ButyricumClostridium butyricum), Lactobacillus sporogenes (Lactobacillus sporogenes), Bacillus subtilis (Bacillus subtilis) And Bacillus polyfermentus(B. polyfermenticus) SCD Etc.

한편, 프로바이오틱 생균제로서 사용되기 위해서는 안전성, 기능성(생존성, 정착성, 서식성, 항미생물제 생성능, 면역 촉진능, 안티제노톡식(antigenotoxic) 활성, 병원성 세균의 억제능), 기술성(관능적 특성, 안정성, 박테리오파지(bacteriophage) 저항성, 제조 과정 중의 생존성), 그리고 GRAS(Generally Recognized As Safe) 미생물로서 큰 장내 생존성 등의 특성이 요구된다.On the other hand, in order to be used as a probiotic probiotic, safety, functionality (survival, fixation, formatting, antimicrobial production ability, immune promoting ability, antigenotoxic activity, inhibitory ability of pathogenic bacteria), technical characteristics (functional characteristics, Properties such as stability, bacteriophage resistance, viability during manufacturing, and large intestinal viability as GRAS (Generally Recognized As Safe) microorganisms are required.

헬리코박터 파이로리(H. pylori)균은 만성위염, 위궤양, 십이지장궤양, 위염발생과 소화성 궤양의 원인균으로서 1982년 와렌(Warren)과 마샬(Marshall)에 의해 보고된 이래 균의 생리적 특성과 병원성 기작 등을 밝히기 위한 많은 연구가 진행되고 있다. 주로 위점막에서 서식하고 있는 헬리코박터 파이로리(H.pylori)균은 날아가는 갈매기 모양인 만곡형 그람음성 세균이며, 크기는 2∼7×0.4∼1.2 ㎛정도이다. 미호기성(microaerophilic)으로 최적 산소분압은 2∼8%이고, 10%의 CO2조건하 37℃에서 배양할 수 있다. 강산성인 위내의 점막에 기생하는 세균이기 때문에 헬리코박터 파이로리(H. pylori)균이 산성에 잘 견디는 균일 것으로 기대하기 쉽지만, 이 균의 최적성장 pH는 7.0∼7.4이며, 이 범위를 벗어난 산성이나 알칼리성 조건에서는 발육하지 않는다(Hazell, S.L., Lee, A. and Hennessy, W.Campylobacter pyloridisand gastritis: Association to an environment of mucus as important factors in colonization of the gastric epithelium. J. Inf. Dis. 153: 658-663 (1986); Steer, H.W. Ultrastructure of cell migration through the gastric epithelium and its relationship to bacteria. J. Clin. Path. 28: 639-646 (1975)).Helicobacter pylori (H. pylori) bacteria chronic gastritis, gastric ulcer, duodenal ulcer, gastritis occurrence and physiological characteristics of the bacteria since reported as a cause of peptic ulcers in 1982 by Warren (Warren) and Marshall (Marshall) and pathogenic mechanisms, etc. Many studies are underway to clarify. Helicobacter pylori ( H. pylori ), which lives mainly in the gastric mucosa, is a flying gram-shaped, gram-negative bacterium that has a size of 2 to 7 × 0.4 to 1.2 ㎛. Microaerophilic, the optimum oxygen partial pressure is 2 to 8% and can be incubated at 37 ° C under 10% CO 2 conditions. It is easy to expect that H. pylori bacteria are acid resistant because they are parasitic bacteria in the gastric mucosa of the strongly acidic stomach, but the optimum growth pH of these bacteria is 7.0-7.4 (Hazell, SL, Lee, A. and Hennessy, W. Campylobacter pyloridis and gastritis: Association to an environment of mucus as important factors in colonization of the gastric epithelium. J. Inf. Dis. 153: 658-663 (1986); Steer, HW Ultrastructure of cell migration through the gastric epithelium and its relationship to bacteria.J. Clin. Path. 28: 639-646 (1975)).

헬리코박터 파이로리(H. pylori) 균의 가장 특징적인 생물학적 성상은 매우 강력한 운동성과 우레아제(urease) 효소 생산능이다. 메틸셀룰로스(Methylcellulose) 인공점액을 이용한 운동성 실험에서 헬리코박터 파이로리(H. pylori) 균은 대장균의 운동성이 저지되는 점도에 비해 20배 이상 높은 점도에서도 운동성을 나타낸다(Hazell, S.L. and Lee, A.Campylobacter pyloridisureas, hydrogen ion back-diffusion and gastric ulcers. Lancet 1: 15-17 (1986)). 이러한 강력한 운동성 때문에 미꾸라지가 진흙 속을 자유로이 다니듯 헬리코박터 파이로리(H. pylori) 균은 살리실레이트(salicylate)나 에탄올(ethanol)같은 화학물 외에는 수소이온 같은 간단한 화합물도 침투하기가 어려운 점액층을 뚫고 들어갈 수도 있고 점액층내에서도 자유롭게 다니면서 위점막 상피세포의 접합부에 부착하여 살아갈 수 있는 것으로 추정된다. 운동성과 관련이 있는 세균 구조물은 편모이다. 이 세균은 단극 또는 양극성 편모가 하나씩 또는 5∼6개의 다발로 구성되어 있다. 특히 다른 세균편모에서는 관찰되지 않는 쉬쓰(sheath)와 터미널 벌브(terminal bulb)라는 독특한 구조물을 가지고 있는데, 이들에 대한 생물학적 성상에 관해서는 아직 밝혀져 있지 않다.The most characteristic biological properties of H. pylori bacteria are very potent motility and the ability to produce urease enzymes. In kinetic experiments using methylcellulose artificial mucus, H. pylori bacteria exhibited motility even at a viscosity 20 times higher than that of E. coli motility (Hazell, SL and Lee, A. Campylobacter pyloridis). ureas, hydrogen ion back-diffusion and gastric ulcers.Lancet 1: 15-17 (1986)). Because of this powerful motility, H. pylori bacteria penetrate the mucous layer, which is difficult to penetrate simple compounds, such as hydrogen ions, in addition to chemicals such as salicylate and ethanol. It is also thought to be able to live freely in the mucous layer and adhere to the junction of gastric mucosal epithelial cells. Bacterial structures associated with motility are flagella. This bacterium is composed of one or five to six bundles of monopolar or bipolar flagella. In particular, they have unique structures called sheaths and terminal bulbs that are not found in other bacterial flagella, and their biological properties are not yet known.

헬리코박터 파이로리(H. pylori) 균의 또 다른 특성 중 하나는 우레아제(urease; urea aminohydrolase) 생산능이 강하다는 것이다(Lee. A. and Hazell, S.L.Campylobacter pyloriin health and disease: An ecological perspective. Microb. Ecol. Health Dis. 1: 1-16 (1988)). 그 생산 능력은 프로테우스(Proteus) 균종의 생산능에 비해 100배 이상이며, 균체 단백질의 약 6% 정도가 우레아제(urease)로 알려져 있다. 우레아제(Urease)는 요소를 암모니아와 이산화탄소로 분해하는 효소이다. 헬리코박터 파이로리(H. pylori) 균은 위점막내에 낮은 농도로 존재하는 요소를 효과적으로 분해할 수 있으며, 이에 따라 생산되는 암모니아가 세균의 주위 환경을 중화시킴으로써 위내의 강염산에 의한 공격에 대해 대처할 수 있는 것으로 알려져 있다.Another characteristic of H. pylori bacteria is its high ability to produce urease (urease; urea aminohydrolase) (Lee.A. and Hazell, SL Campylobacter pylori in health and disease: An ecological perspective.Microb.Ecol Health Dis. 1: 1-16 (1988)). The production capacity is more than 100 times the production capacity of the Proteus strain , and about 6% of the bacterial protein is known as urease (urease). Urease is an enzyme that breaks down urea into ammonia and carbon dioxide. The bacterium H. pylori can effectively break down urea present in the gastric mucosa, and thus the ammonia produced can counteract the attack by strong hydrochloric acid in the stomach by neutralizing the surrounding environment. It is known.

한편, 현재 헬리코박터 파이로리(H. pylori) 균의 치료법으로는 비스머쓰(bismuth)제제와 메트로니아졸(metronidazole)과 함께 아목시실린(amoxicillin) 또는 테트라사이클린(tetracycline)을 병용처리하는 일반적인 3중요법과 비스머쓰(bismuth)제제, 오메프라졸(omeprazole), 테트라사이클린(tetracycline)과 메트로니아졸(metronidazole)을 혼합한 4중요법 등이 헬리코박터 파이로리(H. pylori)에 우수한 치료효과를 나타낸다고 보고되어 있으나 항생제 투여에 의한 내성균주 출현 및 위 내에서의 약물의 침투성과 치료 후에 다시 성장이 억제되어 있던 균이 재증식하는 문제가 있다.On the other hand, the current treatment of H. pylori is a tri-therapy and bismuth in combination with amoxicillin or tetracycline together with bismuth and metronidazole. (bismuth), omeprazole, tetracycline, and tetratherapy combined with metronidazole have been reported to show excellent therapeutic effects on H. pylori . The emergence of resistant strains and the permeability of drugs in the stomach and the growth of bacteria that have been inhibited again after treatment are problematic.

이에 대한 대안으로 헬리코박터 파이로리(H. pylori) 균에 대한 항균활성이 있는 여러 균주를 생균제 형태로 위장에 공급하여 헬리코박터 파이로리 균의 생육을 억제하는 노력이 이뤄지고 있으나, 일부 균들은 장내에서 암을 일으킬 수 있는 위험때문에 생균제로서 쉽게 사용할 수 없는 문제가 있고, 위장 내부의 강산성으로말미암아 내산성이 없는 균주들은 쉽게 사멸하여 효율적으로 헬리코박터 파이로리(Helicobacter pylori) 균의 생육을 억제할 수 없는 문제가 있다.As an alternative, efforts have been made to suppress the growth of Helicobacter pylori by supplying several strains with antimicrobial activity against H. pylori bacteria to the stomach in the form of probiotics, but some bacteria can cause cancer in the intestine. There is a problem that can not be easily used as a probiotic because of the risk, and the acid resistance due to the strong acidity inside the stomach is easily killed and there is a problem that can not effectively inhibit the growth of Helicobacter pylori bacteria.

이에 본 발명은 상기와 같은 문제점으로부터 안출된 것으로, 발암유발의 위험성도 없고, 위액 및 담즙산에 대한 내성이 높아 장내 생존력이 우수하여, 위암 발생 원인균으로 추정되는 헬리코박터 파이로리(Helicobacter pylori) 균에 대해 항균활성이 이 높은 균주를 확보하고 이를 함유하는 생균제로서의 위장병 예방 및치료용 조성물을 제공하는 것을 그 목적으로 한다.Therefore, the present invention has been devised from the above problems, there is no risk of carcinogenesis, high resistance to gastric juice and bile acids, excellent intestinal viability, antibacterial against Helicobacter pylori bacteria suspected to cause gastric cancer It is an object of the present invention to provide a composition for preventing and treating gastrointestinal diseases as a probiotic containing the same, having a high activity strain.

도 1은 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)에 의한 헬리코박터 파이로리(H. pylori) KCTC 2948의 생육 저해를 보여주는 그래프이다.1 is a graph showing the nose Lactobacillus lactis kusu Helicobacter pylori (H. pylori) growth inhibition of KCTC 2948 by (L. lactis) NK24 (KFCC 11315 ).

상기 목적을 달성하기 위하여, 본 발명은 헬리코박터 파이로리(Helicobacter pylori) 균주에 대해 항균효과가 있는 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)를 유효성분으로 함유하는 것을 특징으로 하는 위장병 예방 및 치료용 식품을 제공한다.In order to achieve the above object, the present invention is a gastrointestinal disease prevention, characterized in that it contains L. lactis NK24 (KFCC 11315) having an antimicrobial effect against Helicobacter pylori strains as an active ingredient And food for treatment.

이하, 본 발명에 대해 더욱 상세히 설명한다.Hereinafter, the present invention will be described in more detail.

프로바이오틱 생균으로서 가져야 할 특성 중 하나는 장내 생존력이 높아야 한다는 점이다. 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)는 젓갈에 분리된 균으로서 대조구에서의 108cfu/ml 값에 대해 pH 2.5의 인공위액에서 106cfu/ml의 생존율을 보이고, pH 4.0의 인공담즙산에서는 69.0%, pH 2.5에서는 74.5% 생존율을 보인다. 즉, 본 균주는 인공위액과 인공담즙산에 대해 높은 내성을 보인다.One of the characteristics of probiotic probiotics is the high intestinal viability. L. lactis NK24 (KFCC 11315) is a bacterium isolated from salted fish and shows survival rate of 10 6 cfu / ml in gastric juice of pH 2.5 against 10 8 cfu / ml value in the control. The survival rate was 69.0% for 4.0 artificial bile acids and 74.5% for pH 2.5. That is, the strain shows high resistance to artificial gastric juice and artificial bile acids.

한편, 섭취된 영양분은 위와 장을 통해 흡수되지 않거나 분해되지 않아서 장내에서 이상발효를 일으킬 수 있는데, 프로바이오틱 생균이 생산하는 유익한 효소는 이러한 가능성을 줄일 수 있다.On the other hand, ingested nutrients are not absorbed or decomposed through the stomach and intestine, which can cause abnormal fermentation in the intestine. The beneficial enzymes produced by probiotic probiotics can reduce this possibility.

즉, 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)는 알파-키모트립신(α-chymotrypsin), 베타-갈락토시다제(β-galactosidase) 등의 유익한 효소를 생산하고, 발암효소로 알려진 베타-글루쿠로니다제(β-glucuronidase)와 같은 효소를 생산하지 않았기 때문에 발암유발의 위험성이 없는 것이다.That is, L. lactis NK24 (KFCC 11315) produces beneficial enzymes such as alpha-chymotrypsin and beta-galactosidase, and is a carcinogenic enzyme. There is no risk of carcinogenesis because it does not produce enzymes such as beta-glucuronidase.

또한, 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)는 여러 항생물질 중 니신(nisin), 리팜피신(rifampicin)에 대해서는 저해를 받지 않고, 스트렙토마이신(Streptomycin)에 대해서는 20 ㎍/ml의 농도에서, 젠타마이신(Gentamycin)에 대해서는 10 ㎍/ml 이상의 농도에서 생육저해를 받는다.In addition, L. lactis NK24 (KFCC 11315) is not inhibited by nisin and rifampicin among other antibiotics, and 20 μg / ml of streptomycin by streptomycin. At concentrations, gentamycin is inhibited at concentrations of 10 μg / ml or more.

그람양성균 중 냉동·냉장 제품에서 식중독균으로 주목받고 있는 리시테리아 모노씨토제네스(Listeria monocytogenes) ATCC 15313에 대한 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)의 생육저해 할로(halo)는 40.0 mm이상이고, 진공포장육, 우유 등에서 식중독을 일으키는 그람음성균인 이에르시니아 슈도투베르쿠로시스(Yersinia pseudotuberculosis) ATCC 6902, 이에르시니아 엔테로코리티카(Y. enterocolitica) ATCC 27729, 어패류에서 식중독원인균인 비브리오 파라하에모리티쿠스(Vibrio parahaemolyticus) ATCC 17802 및 비브리오 불니피쿠스(V. vulnificus)에 대한 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)의 생육저해 할로(halo)는 40.0 mm 이상이다. 즉, 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)는 부패세균과 병원성세균에 대해 비교적 넓은 항균효과를 나타낸 것이다.Among the Gram-positive bacteria, halo of L. lactis NK24 (KFCC 11315) against Listeria monocytogenes ATCC 15313, which is attracting attention as food poisoning bacteria in frozen and refrigerated products, More than 40.0 mm, Yersinia pseudotuberculosis ATCC 6902, Y. enterocolitica ATCC 27729, a Gram-negative bacterium that causes food poisoning in vacuum packaged meat, milk, etc. Growth inhibition halo of L. lactis NK24 (KFCC 11315) against Vibrio parahaemolyticus ATCC 17802 and V. vulnificus was greater than 40.0 mm. to be. In other words, L. lactis NK24 (KFCC 11315) shows a relatively wide antibacterial effect against rot bacteria and pathogenic bacteria.

위암 발생 원인균으로 추정되는 헬리코박터 파이로리(H. pylori) KCTC 2948는 기능성 생균제 개발에 있어 중요한 균주인데, 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315) 와 같이 배양하면 1시간 후 헬리코박터 파이로리(H. pylori) KCTC 2948의 존재는 확인할 수 없고, 단지 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)의 균주의 증식만 확인할 수 있다. 따라서, 락토코쿠스 락티스(L.lactis) NK24 (KFCC 11315)의 헬리코박터 파이로리(H. pylori) KCTC 2948에 대한 뚜렷한 항균 효과가 입증된 것이다.Helicobacter pylori ( H. pylori ) KCTC 2948, which is considered to be the cause of gastric cancer, is an important strain in the development of functional probiotics, and incubated with L. lactis NK24 (KFCC 11315) after 1 hour H. pylori ) The presence of KCTC 2948 can not be confirmed, only proliferation of the strain of L. lactis NK24 (KFCC 11315). Therefore, Lactobacillus nose kusu lactis (L.lactis) will demonstrate that the clear antimicrobial effects of the Helicobacter pylori (H. pylori) in NK24 (KFCC 11315) 2948 KCTC.

이에 본 발명은 젓갈에서 분리된 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)의 생균제로서의 용도에 관한 것으로, 위액 및 담즙산에 대한 내성이 높아 장내 생존력이 우수하고, 기존 균주에 비해 병원성 균에 대한 항균활성이 높으며, 베타-갈락토시다제(β-galactosidase)를 생산하므로 유당불내증을 감소시킬 수 있는 효과가 있으므로 매우 유용하고, 이로 말미암아 기능성 젓갈 등의 개발에 사용될 수 있는 것이다.Accordingly, the present invention relates to the use of L. lactis NK24 (KFCC 11315) isolated from salted fish as a probiotic, has high resistance to gastric juice and bile acids, and thus has excellent intestinal viability and pathogenicity compared to existing strains. High antibacterial activity against bacteria, beta-galactosidase (β-galactosidase) produces a very useful because it has the effect of reducing lactose intolerance, which can be used in the development of functional salted fish.

이하, 본 발명의 구성을 하기 실험예 및 실시예를 들어 더욱 상세히 설명하지만 본 발명의 권리범위가 하기 실험 및 실시 예에만 한정되는 것은 아니다.Hereinafter, the configuration of the present invention will be described in more detail with reference to the following Experimental Examples and Examples, but the scope of the present invention is not limited to the following Experiments and Examples.

실험예 1: 락토코쿠스 락티스(Experimental Example 1: Lactococcus lactis ( L. lactisL. lactis ) NK24 (KFCC 11315)에 의한 헬리코박터 파이로리(Helicobacter pylori by NK24 (KFCC 11315) H. pyloriH. pylori )의 생육저해 조사Growth inhibition investigation of

본 실험예에서는 프로바이오틱 생균의 헬리코박터 파이로리(Helicobacter pylori)에 대한 항균 작용을 조사하였다. 헬리코박터 파이로리(Helicobacter pylori)는 헬리코박터 파이로리(H. pylori) KCTC 2948을 사용하였다. 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315) 균주와 헬리코박터 파이로리(H. pylori) KCTC 2948 균주를 혼합배양하고, 인산완충용액으로 세척하였다. 이때, 균주는 1:1 부피비로 혼합하여 10 mL을 제조하고, 헬리코박터 파이로리(H. pylori) KCTC 2948균주의 배양조건인 37℃, 10% CO2배양기에서 과습한 상태를 유지하면서 배양하였다. 한편, 대조구는 헬리코박터 파이로리(H. pylori) KCTC 2948 10 mL을 단독배양하였다.In this experiment we examined the antimicrobial activity of the bacteria Helicobacter pylori of probiotics (Helicobacter pylori). Helicobacter pylori (Helicobacter pylori) was used as the Helicobacter pylori (H. pylori) 2948 KCTC. Lactobacillus nose kusu lactis (L. lactis) NK24 (KFCC 11315 ) was washed with a strain with Helicobacter pylori (H. pylori) mixing KCTC 2948 strain culture and a phosphate buffer solution. At this time, the strain was mixed in a 1: 1 volume ratio to prepare a 10 mL, and cultured while maintaining a humid state in 37 ℃, 10% CO 2 incubator, the culture conditions of Helicobacter pylori ( H. pylori ) KCTC 2948 strain. On the other hand, the control group was incubated with 10 mL of H. pylori KCTC 2948 alone.

접종 후 1시간이 지났을 때, 헬리코박터 파이로리(H. pylori) KCTC 2948의 존재는 확인할 수 없었고, 단지 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315) 균주의 증식만 확인할 수 있었다(도 1).One hour after inoculation, the presence of H. pylori KCTC 2948 could not be confirmed, and only the proliferation of L. lactis NK24 (KFCC 11315) strains could be confirmed (FIG. 1). ).

이상의 결과에 의해서, 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)의 헬리코박터 파이로리(H. pylori) KCTC 2948에 대한 뚜렷한 항균 효과가 입증된 것이다.By the above results, the Lactobacillus nose kusu lactis (L. lactis) will demonstrate that the clear antimicrobial effects of the Helicobacter pylori (H. pylori) KCTC 2948 in NK24 (KFCC 11315).

실험예 2: 락토코쿠스 락티스(Experimental Example 2: Lactococcus lactis ( L. lactisL. lactis ) NK24 (KFCC 11315)의 인공위액에 대한 내성 조사Investigation of Resistance to Artificial Gastric Fluid of NK24 (KFCC 11315)

본 실험예에서는 프로바이오틱 생균의 인공위액에 대한 내성을 조사하였다. 식품용 프로바이오틱 생균으로서 가져야 할 특성이 여러 가지가 있는데, 이 중에서 가장 중요한 특성 중 하나가 장내에서의 생존력이 높아야 한다는 점이다. 구강을 통하여 섭취되는 균은 위와 십이지장을 통해 생존하여 최종 목적 부위인 장에 도달하기 때문에 프로바이오틱 생균제로서의 기능을 발휘하기 위해서는 강산성의 위액을 통과하여야 한다.In this experimental example, the resistance of probiotic live bacteria to artificial gastric juice was investigated. Probiotic probiotic for food has a number of characteristics, one of the most important of these is the high viability in the intestines. Bacteria ingested through the oral cavity survive through the stomach and duodenum and reach the final target site of the intestine, so they must pass strong acidic gastric juice to function as a probiotic probiotic.

인공위액은 'Kobayashi'의 방법(Kobayashi, Y., Tohyama, K. and Terashima,T. Tolerance of the multuple antibiotic resistant strain, L. casei PSR 3002, to artificial digestive fluids. Jpn. J. Microbiol. 29: 691-697 (1974))에 따라 1 N HCl을 사용하여 pH 4.0, 2.5로 조정한 'MRS broth' 에 1%(v/v) 펩신을 첨가하여 10 mL을 제조하여 사용하였다.Artificial gastric fluid was obtained from the method of Kobayashi (Kobayashi, Y., Tohyama, K. and Terashima, T. Tolerance of the multuple antibiotic resistant strain, L. casei PSR 3002, to artificial digestive fluids.Jpn. J. Microbiol. 29: 691-697 (1974)) was used to prepare 10 mL by adding 1% (v / v) pepsin to 'MRS broth' adjusted to pH 4.0, 2.5 using 1 N HCl.

락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)는 그 배양액을 'eppendorf tube'에 1 ml 넣은 후 원심분리(6,000 rpm, 10 min)한 후 균체 만을 회수하여 사용하였다. L. lactis NK24 (KFCC 11315) was used to collect the cells after centrifugation (6,000 rpm, 10 min) after putting 1 ml of the culture solution into the 'eppendorf tube'.

회수된 균체는 인공위액에 첨가하여 37℃, 2시간동안 배양하여 총균수를 측정하였다. 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)는 대조구의 108cfu/ml의 값에 대해 pH 2.5에서 106cfu/ml의 생존율을 보였다.The recovered cells were added to artificial gastric juice and incubated at 37 ° C. for 2 hours to determine the total bacterial count. L. lactis NK24 (KFCC 11315) showed a survival rate of 10 6 cfu / ml at pH 2.5 for a value of 10 8 cfu / ml of the control.

락토코쿠스 락티스(Lactococcus lactis) NK24 (KFCC 11315)의 인공 위액에서 생존율Survival rate in artificial gastric juice of Lactococcus lactis NK24 (KFCC 11315) 대조구 (CFU/ml)Control (CFU / ml) pH 4.0 (CFU/ml)pH 4.0 (CFU / ml) pH 2.5 (CFU/ml)pH 2.5 (CFU / ml) 3.0×108 3.0 × 10 8 2.0×108(53.3%)2.0 × 10 8 (53.3%) 4.0×106(1.3%)4.0 × 10 6 (1.3%)

실험예 3: 락토코쿠스 락티스(Experimental Example 3: Lactococcus lactis ( L. lactisL. lactis ) NK24 (KFCC 11315)의 인공 담즙산에 대한 내성 조사Investigation of NK24 (KFCC 11315) Resistance to Artificial Bile Acids

본 실험예에서는 인공 담즙산에 대한 내성을 조사하였다. 섭취된 락토코쿠스 락티스(Lactococcus lactis) NK24 (KFCC 11315) 프로바이오틱 생균이 장에 도달하기 위해서 위를 거쳐 췌장과 십이지장을 통과하게 된다. 이 부위에서 분비되는 담즙액에 대한 내성 또한 프로바이오틱 생균이 가져야 할 중요한 특성이다.In this experimental example, resistance to artificial bile acids was investigated. Ingested Lactococcus lactis NK24 (KFCC 11315) probiotic probiotics pass through the stomach and pancreas and duodenum to reach the intestine. Resistance to the bile secreted from this site is also an important property of probiotic probiotics.

인공담즙액은 멸균된 각각의 브로쓰(broth)에 멸균한 10%(w/v) 'oxgall' 용액을 1%(v/v) 첨가하여 10 mL을 제조하여 사용하였다. 인공위액을 거친 배양액 1 mL을 인공담즙액에 첨가하였다. 이후 37℃, 24시간 동안 배양하여 총균수를 측정하였다. 인공담즙에 대한 내성을 실험한 결과 pH 4.0에서는 69.0%, pH 2.5에서는 74.5% 생존율을 보였다.The artificial bile solution was used to prepare 10 mL by adding 1% (v / v) of sterilized 10% (w / v) 'oxgall' solution to each sterilized broth. 1 mL of culture solution subjected to artificial gastric juice was added to the artificial bile. After incubation for 24 hours at 37 ℃, the total number of bacteria was measured. The resistance to artificial bile was 69.0% for pH 4.0 and 74.5% for pH 2.5.

따라서, 본 균주는 인공위액과 인공담즙산에 대해 내성이 있는 것이다.Therefore, this strain is resistant to artificial gastric juice and artificial bile acid.

락토코쿠스 락티스(Lactococcus lactis) NK24 (KFCC 11315)의 인공 위액을 거친 후, 인공 담즙산에서의 생존율Survival rate in artificial bile acids after artificial gastric juice of Lactococcus lactis NK24 (KFCC 11315) pH 4.0pH 4.0 pH 2.0pH 2.0 대조구(CFU/ml)Control (CFU / ml) 담즙산(CFU/ml)Bile Acids (CFU / ml) 대조구(CFU/ml)Control (CFU / ml) 담즙산(CFU/ml)Bile Acids (CFU / ml) 3.9×109 3.9 × 10 9 2.7×109(69.0%)2.7 × 10 9 (69.0%) 2.5×109 2.5 × 10 9 1.8×109(74.5%)1.8 × 10 9 (74.5%)

실험예 4: 락토코쿠스 락티스(Experimental Example 4: Lactococcus lactis ( L. lactisL. lactis ) NK24 (KFCC 11315)의 효소활성 측정) Determination of Enzyme Activity of NK24 (KFCC 11315)

본 실험예에서는 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315) 프로바이오틱 생균의 효소활성을 조사하였다. 프로바이오틱 생균이 가져야 할 특성 중에 이들 균이 생성하는 효소 또한 중요한 부분을 차지하고 있기 때문이다.In this experimental example, the enzymatic activity of L. lactis NK24 (KFCC 11315) probiotic probiotic was investigated. This is because the enzymes produced by these bacteria are also an important part of the probiotic probiotics.

락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)의 배양액 10 ml을 원심분리하여 균체만을 회수하여 멸균수로서 2회 반복하여 세척하고 멸균수 10 ml로 현탁하였다. 이 현탁액 100 μl를 API ZYM(BioMeriux Co.)에 접종하여 어두운 곳에 37℃, 4시간동안 배양하였다. 표현활성 증가와 용해를 돕기 위해 ZYM A, B시약을 각각의 큐플에 한 방울씩 떨어뜨린 후 밝은 곳에서 약 5분간 반응시킨 후 색깔의 변화를 관찰하여 효소활성을 측정하였다.10 ml of the culture solution of L. lactis NK24 (KFCC 11315) was centrifuged to recover only the cells, washed twice with sterile water, and suspended in 10 ml of sterile water. 100 μl of this suspension was inoculated in API ZYM (BioMeriux Co.) and incubated for 4 hours at 37 ° C. in the dark. ZYM A and B reagents were dropped one drop into each chapel to help increase expression and dissolution, and then reacted for 5 minutes in a bright place.

그 결과, 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)는 알파-키모트립신(α-chymotrypsin), 베타-갈락토시다제(β-galctosidase) 등의 유익한 효소를 생산하였고, 발암효소로 알려진 베타-글루쿠로니다제(β-glucuronidase)와 같은 효소를 생산하지 않기 때문에 발암유발의 위험성이 없다는 것을 알 수 있었다.As a result, L. lactis NK24 (KFCC 11315) produced beneficial enzymes such as alpha-chymotrypsin and beta-galctosidase, and carcinase. Because it does not produce enzymes such as beta-glucuronidase, it is known that there is no risk of carcinogenesis.

API ZYM 키트에 의한 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)의 효소활성 측정Determination of Enzyme Activity of L. lactis NK24 (KFCC 11315) by API ZYM Kit 효소enzyme 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315) L. lactis NK24 (KFCC 11315) 대조구Control 00 Alkaline phosphataseAlkaline phosphatase 44 Esterase(C4)Esterase (C 4 ) 00 Esterase Lipase(C8)Esterase Lipase (C 8 ) 44 Lipase(C14)Lipase (C 14 ) 00 Leucine acrylamidaseLeucine acrylamidase 55 Valine arylamidaseValine arylamidase 00 Crystine arylamidaseCrystine arylamidase 22 TrypsinTrypsin 00 α-Chymotrypsinα-Chymotrypsin 33 Acid phosphateAcid phosphate 55 Naphthol-AS-BI-phosphohydrolaseNaphthol-AS-BI-phosphohydrolase 55 α-Galactosidaseα-Galactosidase 00 β-Galactosidaseβ-Galactosidase 44 β-Glucuronidaseβ-Glucuronidase 00 α-Glucosidaseα-Glucosidase 22 β-Glucosidaseβ-Glucosidase 1One N-Acetyl-β-glucosaminidaseN-Acetyl-β-glucosaminidase 00 α-Mannosidaseα-Mannosidase 00 β-Fucosidaseβ-Fucosidase 00 0; 0 nmol, 1; 5 nmol, 2; 10 nmol, 3; 20 nmol, 4; 30 nmol, 5; 40 nmol.0; 0 nmol, 1; 5 nmol, 2; 10 nmol, 3; 20 nmol, 4; 30 nmol, 5; 40 nmol.

실험예 5: 락토코쿠스 락티스(Experimental Example 5: Lactococcus lactis ( L. lactisL. lactis ) NK24 (KFCC 11315)의 항생물질에 대한 내성 조사Investigation of resistance to antibiotics of NK24 (KFCC 11315)

본 실험예에서는 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315) 프로바이오틱 생균의 항생물질에 대한 내성을 조사하였다. 항생물질 내성을 측정하기 위해서 'paper disc' 방법을 사용하였다. 즉, 균주를 최적의 온도에서 12시간 배양하고, 각각의 'soft agar (0.7%)'에 배양액을 100 μl로 접종하여 아가 평판에 오버레이(overlay)하였다. 그 후 오버레이된 평판에서 표 4에 기재된 조건에 의하여 항생물질에 대한 내성 측정하였다.In this experiment, L. lactis NK24 (KFCC 11315) probiotic probiotic resistance to antibiotics was investigated. The paper disc method was used to measure antibiotic resistance. That is, the strains were incubated at the optimal temperature for 12 hours, and 100 μl of the culture solution was inoculated into each 'soft agar (0.7%)' to overlay the agar plate. The resistance to antibiotics was then measured by the conditions described in Table 4 on the overlayed plates.

락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)의 항생물질에 대한 내성조사Investigation on the Antibiotic Resistance of L. lactis NK24 (KFCC 11315) 항생물질 (㎍/ml)Antibiotic (µg / ml) 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315) L. lactis NK24 (KFCC 11315) NisinNisin 0255010002550100 ++++++++ GentamycinGentamycin 051020051020 ++--++- RifampicinRifampicin 051020051020 ++++++++ ChloramphenicolChloramphenicol 051020051020 +---+ --- RoxithromycinRoxithromycin 051020051020 +---+ --- StreptomyinStreptomyin 051020051020 +++-+++- ErythromycinErythromycin 051020051020 +---+ --- +; growth, -; no growth.+; growth,-; no growth.

그 결과, 니신(nisin), 리팜피신(rifampicin)에 대해서는 저해를 받지 않았고, 스트렙토마이신(Streptomycin)에 대해서는 20 ㎍/ml의 농도에서,젠타마이신(Gentamycin)에 대해서는 10 ㎍/ml 이상의 농도에서 생육저해를 받음을 확인하였다.As a result, it was not inhibited by nisin and rifampicin, but inhibited growth at a concentration of 20 µg / ml for streptomycin and 10 µg / ml or higher for gentamycin. Confirmed receiving.

실험예 6: 락토코쿠스 락티스(Experimental Example 6: Lactococcus lactis ( L. lactisL. lactis ) NK24 (KFCC 11315)의 항균활성 조사) Antibacterial Activity of NK24 (KFCC 11315)

본 실험예에서는 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315) 프로바이오틱 생균의 항균활성을 측정하였다. 항균활성을 측정하기 위해 'modified deferred method'를 사용하였다. 즉, 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)는 살균된 MRS broth 10 ml에 각각 한 백금이 접종하여 최적 배양온도에서 12시간 배양한 후 건조된 'MRS agar' 평판 표면에 배양액 5 μl을 스팟하고 37℃에서 24시간 배양하였다.In this experimental example, the antimicrobial activity of L. lactis NK24 (KFCC 11315) probiotic probiotic was measured. The 'modified deferred method' was used to measure the antimicrobial activity. In other words, L. lactis NK24 (KFCC 11315) was inoculated with 10 ml of sterile MRS broth, incubated for 12 hours at the optimum culture temperature, and then cultured on the dried 'MRS agar' plate surface. 5 μl were spotted and incubated at 37 ° C. for 24 hours.

피검균주들은 최적배지와 최적온도에서 12시간 배양하였다. 이 배양액 100 ㎕를 각각 0.75% 'soft agar'에 접종하여 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)가 자란 평판에 오버레이(overlay)하고 최적온도에서 24시간 배양하여 활성을 측정하였다.Test strains were incubated for 12 hours at the optimal medium and temperature. 100 μl of this culture solution was inoculated in 0.75% 'soft agar', respectively, and overlayed onto a plate grown with L. lactis NK24 (KFCC 11315) and incubated at an optimal temperature for 24 hours to measure activity. .

락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)는 그람양성균 중 리스테리아 모노씨토제네스(Listeria monocytogenes) ATCC 15313에 대해 생육저해 할로(halo)로 40.0 mm이상을 나타내었고, 그람음성균인 이에르시니아 슈도투베르쿠로시스(Yersinia pseudotuberculosis) ATCC 6902, 이에르시니아 엔테로코리티카(Y. enterocolitica) ATCC 277929, 비브리오 파라하에모리티쿠스(V. parahaemolyticus) ATCC 17802 및 비브리오 불니피쿠스(V. vulnificus)에 대해서는 생육저해 할로로 40.0 mm이상을 나타내었다(표 5). L. lactis NK24 (KFCC 11315) showed more than 40.0 mm of halo against Listeria monocytogenes ATCC 15313 among Gram-positive bacteria. Yersinia pseudotuberculosis ATCC 6902, Y. enterocolitica ATCC 277929, V. parahaemolyticus ATCC 17802, and V. vulnificus ) Showed more than 40.0 mm growth halo (Table 5).

락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)의 부패성 및 병원성균에 대한 항균활성Antimicrobial Activity of L. lactis NK24 (KFCC 11315) against Decay and Pathogenic Bacteria 균주Strain 저해환직경(Inhibition zoneDiameter)(mm)Inhibition zone diameter (mm) 균주Strain 저해환직경(Inhibition zoneDiameter)(mm)Inhibition zone diameter (mm) Gram positive bacteriaGram positive bacteria Escherichia coliKCCM 32396 Escherichia coli KCCM 32396 18.518.5 Bacillus cereusATCC 11778 Bacillus cereus ATCC 11778 16.016.0 Escherichia coliJM 109 Escherichia coli JM 109 17.017.0 Bacillus cereusBacillus cereus 17.017.0 Escherichia coliO157:H7 Escherichia coli O157: H7 19.519.5 Bacillus pumilisBacillus pumilis 26.026.0 Pseudomonas aeruginosaATCC 15442 Pseudomonas aeruginosa ATCC 15442 31.531.5 Bacillus subtilisIFO 12113 Bacillus subtilis IFO 12113 23.023.0 Pseudomonas fluorescensPseudomonas fluorescens 12.012.0 Bacillus subtilisATCC 19433 Bacillus subtilis ATCC 19433 24.024.0 Pseudomonas putidaPseudomonas putida >40.0> 40.0 Clostridium perfringensATCC 3624 Clostridium perfringens ATCC 3624 32.032.0 Salmonella typhiSalmonella typhi 16.016.0 Enterococcus faecalisATCC 19433 Enterococcus faecalis ATCC 19433 6.56.5 Salmonella paratyphiA Salmonella paratyphi A 21.521.5 Lactobacillus delbruekiiATCC 4797 Lactobacillus delbruekii ATCC 4797 24.524.5 Salmonella typhimuriumSalmonella typhimurium 30.030.0 Listeria monocytogenesATCC 15313 Listeria monocytogenes ATCC 15313 >40.0> 40.0 Salmonella enteritidisSalmonella enteritidis 16.016.0 Listeria ivanoviiATCC 19119 Listeria ivanovii ATCC 19119 12.012.0 Shigella flexneriShigella flexneri 20.020.0 Micrococcus flavusATCC 10240 Micrococcus flavus ATCC 10240 >40.0> 40.0 Shigella boydiiShigella boydii 32.032.0 Pediococcus acidilacticiKCTC 1626 Pediococcus acidilactici KCTC 1626 25.025.0 Shigella sonneiShigella sonnei 21.521.5 Staphylococcus aureusATCC 25923 Staphylococcus aureus ATCC 25923 24.024.0 Sphingomonas paucimobilisSphingomonas paucimobilis >40.0> 40.0 Staphylococcus aureusKCCM 32959 Staphylococcus aureus KCCM 32959 34.034.0 Vibrio parahaemolyticusATCC 17802 Vibrio parahaemolyticus ATCC 17802 >40.0> 40.0 Staphylococcus epidermidisATCC12228 Staphylococcus epidermidis ATCC12228 21.021.0 Vibrio vulnificusVibrio vulnificus >40.0> 40.0 Gram negative bacteriaGram negative bacteria Vibrio choleraO139 Vibrio cholera O139 24.524.5 Chryseomonas luteolaChryseomonas luteola 00 Yersinia enterocoliticaATCC 27729 Yersinia enterocolitica ATCC 27729 >40.0> 40.0 Escherichia coliATCC 8739 Escherichia coli ATCC 8739 20.020.0 Yersinia pseudotuberculosisATCC6902 Yersinia pseudotuberculosis ATCC6902 >40.0> 40.0 Escherichia coliATCC 29522 Escherichia coli ATCC 29522 17.517.5 Xanthomonas maltophilaXanthomonas maltophila 17.017.0

따라서, 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)는 부패균과 병원성균에 대해 비교적 넓은 항균효과를 나타낸다고 할 수 있다.Therefore, L. lactis NK24 (KFCC 11315) can be said to have a relatively wide antibacterial effect against rot and pathogenic bacteria.

위와 같은 결과에 의해 식중독 원인균에 의해 감염이 되었을 경우 장내에 상주하는 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315) 프로바이오틱 균주들에 의해 어느 정도 억제될 수 있을 것이라 판단되었다.Based on the above results, it was determined that L. lactis NK24 (KFCC 11315) probiotic strains resident in the intestine may be inhibited to some extent when infected by food poisoning bacteria.

한편, 본 발명의 바람직한 실시예는 하기와 같다.On the other hand, preferred embodiments of the present invention are as follows.

실시예 1: 락토코쿠스 락티스(Example 1 Lactococcus lactis L. lactisL. lactis ) NK24 (KFCC 11315)을 포함하는 위장병 예방 및 치료제의 제조) Preparation of gastrointestinal disease prevention and treatment containing NK24 (KFCC 11315)

락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)이 함유된 위장약은 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)을 통상의 진공동결건조기에 투입하여 동결건조한다음 캡슐화시켜 제조하였다.Lactobacillus nose kusu lactis (L. lactis) NK24 (KFCC 11315 ) containing the Gastrointestinal drug is prepared Lactobacillus nose kusu lactis (L. lactis) by putting the NK24 (KFCC 11315) in a conventional vacuum freeze dryer, freeze-dried encapsulated by It was.

실시예 2: 락토코쿠스 락티스(Example 2 Lactococcus lactis L. lactisL. lactis ) NK24 (KFCC 11315)를 포함하는 건조효모가공식품의 제조) Preparation of Dried Yeast Processed Foods Containing NK24 (KFCC 11315)

아포형태로 공급된 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315) 14 g을 맥주효모 80 g, 솔비톨 5 g, 해조칼슘 1 g에 첨가하여 건조효모가공식품을 제조하였다.14 g of L. lactis NK24 (KFCC 11315) supplied in apo form was added to 80 g of beer yeast, 5 g of sorbitol, and 1 g of seaweed calcium to prepare a dried yeast processed food.

실시예 3: 락토코쿠스 락티스(Example 3: Lactococcus lactis L. lactisL. lactis ) NK24 (KFCC 11315)를 포함하는 선식의 제조) Preparation of Wires Containing NK24 (KFCC 11315)

찹쌀 15 g, 보리쌀 20 g, 멥쌀 15 g, 현미 10 g, 흰콩 5 g, 수수 1 g, 조 1 g, 밀 1 g, 메밀 1 g, 옥수수 5 g, 밤 1 g, 잣 1 g, 호두 1 g, 당근 1 g, 시금치 1 g, 마 1 g, 연자육 1 g, 홍화씨 1 g, 둥글레 1 g, 다시마 2 g, 해태 1 g, 뽕잎 3 g, 누에 3 g, 동충하초 3 g을 함유하는 조성물에 아포형태로 공급된 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315) 5 g을 첨가하여 선식을 제조하였다.15 g of glutinous rice, 20 g of barley rice, 15 g of brown rice, 10 g of brown rice, 5 g of white beans, 1 g of crude, 1 g of wheat, wheat 1 g, buckwheat 1 g, corn 5 g, chestnut 1 g, pine nuts 1 g, walnut 1 g, carrot 1 g, spinach 1 g, hemp 1 g, lotus root 1 g, safflower seed 1 g, roundle 1 g, kelp 2 g, Haitai 1 g, mulberry leaf 3 g, silkworm 3 g, cordyceps 3 g The wire was prepared by adding 5 g of L. lactis NK24 (KFCC 11315) supplied in apoform form.

실시예 4: 위장병 예방 및 치료용 조성물의 일 예로, 락토코쿠스 락티스(Example 4 As an example of a composition for preventing and treating gastrointestinal diseases, Lactococcus lactis ( L. lactisL. lactis ) NK24 (KFCC 11315)를 함유한 꽁치젓갈의 제조) Preparation of Saury Salted Fish with NK24 (KFCC 11315)

꽁치 약 78%(v/v)에 소금 약 22%(v/v) 정도의 비율로 상호 혼합하여 지하에 설치되는 저장탱크, 즉 발효탱크에 1 L 투입한 후 발효탱크의 온도를 약 14℃ 정도가 되도록 하여 약 5개월 정도 발효시킨 후, 발효된 꽁치젓갈에서 침출된 침출액 약 25%(v/v) 정도를 제거한 후 고형분 꽁치 덩어리를 약 0.1mm 정도의 입자크기로 분쇄하였다. 이어 여기에 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)를 꽁치젓갈 전체부피 대비 0.05%(v/v) 첨가하여 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315) 함유 기능성 꽁치젓갈을 제조하였다.1% of fermentation tank after mixing 1% into the storage tank installed underground, ie fermentation tank, mixed with each other at about 78% (v / v) and about 22% (v / v) salt. After about 5 months of fermentation, about 25% (v / v) of the leaching liquor from the fermented saury was removed, and the solid saury chunk was crushed to a particle size of about 0.1 mm. Subsequently, L. lactis NK24 (KFCC 11315) was added to 0.05% (v / v) of the total volume of saury, and it contained L. lactis NK24 (KFCC 11315). Saury salted fish was prepared.

실시예 5: 위장병 예방 및 치료용 조성물의 일 예로, 락토코쿠스 락티스(Example 5 As an example of a composition for preventing and treating gastrointestinal diseases, Lactococcus lactis ( L. lactisL. lactis ) NK24 (KFCC 11315)를 함유한 멸치액젓의 제조) Preparation of Anchovy Sauce Sauce Containing NK24 (KFCC 11315)

멸치 약 78%(v/v)에 소금 약 22%(v/v) 정도의 비율로 상호 혼합하여 발효탱크에 1 L 첨가한 후, 온도를 약 14℃ 정도가 되도록 하여 약 12개월 정도 발효시켜서 침출된 멸치액젓을 제조하였다. 이어 여기에 여기에 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315)를 멸치액젓 전체부피 대비 0.05%(v/v) 첨가하여 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315) 함유 기능성 멸치액젓을 제조하였다.Anchovy about 78% (v / v) mixed with salt at a rate of about 22% (v / v), added 1 liter to the fermentation tank, and then fermented for about 12 months at a temperature of about 14 ℃. A leached anchovy sauce was prepared. Next, L. lactis NK24 (KFCC 11315) was added to 0.05% (v / v) of the total volume of anchovy fish sauce and L. lactis NK24 (KFCC 11315). Containing functional anchovy sauce was prepared.

이상 발명의 상세한 설명에서 확인할 수 있는 바와 같이, 본 발명의 락토코쿠스 락티스(L. lactis) NK24 (KFCC 11315) 생균이 함유된 조성물은 헬리코박터 파이로리(Helicobacter pylori) 균에 대해 항균활성이 있을 뿐만 아니라, 위액 및 담즙산에 대한 내성이 높아 장내 생존력이 우수하고, 기존 균주에 비해 병원성 균에 대한 항균활성이 높으며, 베타-갈락토시다제(β-galactosidase)를 생산하므로 유당불내증을 감소시키는 우수한 효과가 있으므로 식품, 의약품 등의 용도로 사용될 수 있는 매우 유용한 발명인 것이다.As can be seen in the detailed description of the invention, the composition containing L. lactis NK24 (KFCC 11315) probiotic of the present invention has an antimicrobial activity against Helicobacter pylori bacteria. In addition, it has excellent resistance to gastric juice and bile acids, and therefore has excellent intestinal viability, high antimicrobial activity against pathogenic bacteria compared to existing strains, and produces beta-galactosidase, thereby reducing lactose intolerance. Since there is a very useful invention that can be used for food, pharmaceuticals and the like.

Claims (1)

락토코쿠스 락티스(Lactococcus lactis) NK24 (KFCC 11315)를 유효성분으로 함유하는 것을 특징으로 하는 위장병 예방 및 치료용 조성물. Lactococcus lactis ( Lactococcus lactis ) A composition for preventing and treating gastrointestinal diseases, characterized by containing NK24 (KFCC 11315) as an active ingredient.
KR10-2002-0072758A 2002-11-21 2002-11-21 Composition containing Lactococcus lactis NK24 (KFCC 11315) for stomach disorder KR100518263B1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
KR10-2002-0072758A KR100518263B1 (en) 2002-11-21 2002-11-21 Composition containing Lactococcus lactis NK24 (KFCC 11315) for stomach disorder

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
KR10-2002-0072758A KR100518263B1 (en) 2002-11-21 2002-11-21 Composition containing Lactococcus lactis NK24 (KFCC 11315) for stomach disorder

Publications (2)

Publication Number Publication Date
KR20040044300A true KR20040044300A (en) 2004-05-28
KR100518263B1 KR100518263B1 (en) 2005-10-04

Family

ID=37340555

Family Applications (1)

Application Number Title Priority Date Filing Date
KR10-2002-0072758A KR100518263B1 (en) 2002-11-21 2002-11-21 Composition containing Lactococcus lactis NK24 (KFCC 11315) for stomach disorder

Country Status (1)

Country Link
KR (1) KR100518263B1 (en)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100837857B1 (en) * 2006-12-14 2008-06-13 한국식품연구원 Bacillus sp., isolated from meolchi-jeot, having fibrinolytic, immunostimulating activities and jeot-kal products containing thereof
CN100448981C (en) * 2005-09-16 2009-01-07 新疆农业科学院微生物应用研究所 Beta-galactosidase strain, beta-galactosidase and its production process
US9375444B2 (en) 2008-08-16 2016-06-28 Protectimmun Gmbh Composition for prevention and treatment of allergic and/or inflammatory diseases
US9950017B2 (en) 2007-03-02 2018-04-24 Forschungszentrum Borstel Pharmaceutical composition for protection from allergies and inflammatory disorders
CN115386523A (en) * 2022-09-30 2022-11-25 北京诺安百汇医药科技有限公司 Lactococcus lactis and application thereof in resisting helicobacter pylori infection

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5804549A (en) * 1996-01-05 1998-09-08 Ambi Inc. Compositions with activity against helicobacter
AU4059999A (en) * 1998-06-05 1999-12-30 Wakamoto Pharmaceutical Co., Ltd. Lactic acid bacterium-containing compositions, drugs and foods
KR100324441B1 (en) * 1999-02-08 2002-02-27 이은선 Food for preventing gastritis, gastric and duodenal ulcers
KR100375342B1 (en) * 2000-02-08 2003-03-08 주식회사 풀 무 원 A natural antimicrobial agents for the improvement of gastrointestinal disease.
KR100357668B1 (en) * 2000-02-19 2002-10-18 주식회사 한국야쿠르트 Lactobacillus acidophilus HY 2177, Lactobacillus casei HY 2743 having antibiotic activation for Helicobacter pylori and Lactobacillus product thereof
KR100413335B1 (en) * 2001-05-30 2003-12-31 주식회사풀무원 Lactococcus lactis BH5(KCCM-10275) which is produced antimicrobial peptide

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100448981C (en) * 2005-09-16 2009-01-07 新疆农业科学院微生物应用研究所 Beta-galactosidase strain, beta-galactosidase and its production process
KR100837857B1 (en) * 2006-12-14 2008-06-13 한국식품연구원 Bacillus sp., isolated from meolchi-jeot, having fibrinolytic, immunostimulating activities and jeot-kal products containing thereof
US9950017B2 (en) 2007-03-02 2018-04-24 Forschungszentrum Borstel Pharmaceutical composition for protection from allergies and inflammatory disorders
US9375444B2 (en) 2008-08-16 2016-06-28 Protectimmun Gmbh Composition for prevention and treatment of allergic and/or inflammatory diseases
CN115386523A (en) * 2022-09-30 2022-11-25 北京诺安百汇医药科技有限公司 Lactococcus lactis and application thereof in resisting helicobacter pylori infection
CN115386523B (en) * 2022-09-30 2024-01-30 北京诺安百汇医药科技有限公司 Lactococcus lactis and application thereof in resisting helicobacter pylori infection

Also Published As

Publication number Publication date
KR100518263B1 (en) 2005-10-04

Similar Documents

Publication Publication Date Title
US5951977A (en) Competitive exclusion culture for swine
US7192581B2 (en) Lactobacillus reuteri useful as probiotics
Johansson et al. Administration of different Lactobacillus strains in fermented oatmeal soup: in vivo colonization of human intestinal mucosa and effect on the indigenous flora
EP0554418B1 (en) Intestine colonizing lactobacilli
US7927584B2 (en) Lactic bacteria useful as probiotics
Ljungh et al. Isolation, selection and characteristics of Lactobacillus paracasei subsp. paracasei F19
CN110373368B (en) Bifidobacterium longum strain ZJ1 and application thereof
US6841149B1 (en) Probiotic mixture intended for monogastric animals to control intestinal flora populations
KR100443254B1 (en) Lactobacillus plantarum having acid-, bile acid- and antibiotic-resistance
KR100518263B1 (en) Composition containing Lactococcus lactis NK24 (KFCC 11315) for stomach disorder
KR101098946B1 (en) A compound for feed additive comprising novel Lactobacillus salivarius G1-1
KR20030064030A (en) Novel probiotic strain isolated from Korean feces having gastric juice-resistance, bile acid-resistance and its use
KR20220004865A (en) Lactobacillus brevis SRCM101607 strain having probiotics-related enzyme secretion activity, antioxidant activity, Bile salt hydrolysis, antimicrobial activity, and not producing harmful enzyme and harmful metabolite and uses thereof
US6524574B1 (en) Probiotic mixture intended for monogastric animals to control intestinal flora populations
KR100803532B1 (en) - DF20KCTC10942BP Lactobacillus salivarius sp. salivarius DF20 having been Acid-tolerant Bile-tolerant Antibacterial activity and possesed Alpha-galactosidase
KR100424659B1 (en) Novel probiotic strain having alcohol-resistance and its use
KR101201337B1 (en) A feed additive containig novel Lactobacillus spp. complex
KR100533677B1 (en) Feed additive for fish-breeding containing Lactobacillus plantarum as effective ingradient
Lee et al. Potential probiotic properties of Lactococcus lactis NK34 isolated from jeotgal
KR100320031B1 (en) Bispan mutant with enhanced bacteriocin productivity and process for praparation bacteriocin using the same
KR101797139B1 (en) Novel Lactobacillus gallinarum strain and uses thereof
KR100376954B1 (en) Lactobacillus colonized in the intestine
KR20030075716A (en) Composition containing anti-Helicobacter pylori Bacillus polyfermenticus SCD (KCCM 10104) or polyfermenticin SCD
KR20240001770A (en) Lactobacillus brevis SRCM 101610 strain having β-galactosidase secretion activity, bile salt hydrolytic activity and probiotics property and uses thereof
KR20090057154A (en) Lactobacillus sp. isolated from chicken feces

Legal Events

Date Code Title Description
A201 Request for examination
E902 Notification of reason for refusal
E701 Decision to grant or registration of patent right
GRNT Written decision to grant
FPAY Annual fee payment

Payment date: 20120914

Year of fee payment: 8

FPAY Annual fee payment

Payment date: 20130912

Year of fee payment: 9

LAPS Lapse due to unpaid annual fee