KR101649476B1 - Growth inhibitor against bacillus amyloliquefaciens KNU-1 (KCTC18343P) or culture medium thereof - Google Patents
Growth inhibitor against bacillus amyloliquefaciens KNU-1 (KCTC18343P) or culture medium thereof Download PDFInfo
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- KR101649476B1 KR101649476B1 KR1020150033361A KR20150033361A KR101649476B1 KR 101649476 B1 KR101649476 B1 KR 101649476B1 KR 1020150033361 A KR1020150033361 A KR 1020150033361A KR 20150033361 A KR20150033361 A KR 20150033361A KR 101649476 B1 KR101649476 B1 KR 101649476B1
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Abstract
Description
본 발명은 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 또는 이의 배양액을 포함하는 생육저해제에 관한 것이다.
The present invention is Bacillus amyl Lowry query Pacific Enschede (Bacillus amyloliquefaciens KNU-1 (KCTC18343P) or a culture thereof.
자연계에 분포해 있는 유해 미생물은 감염 피해를 불러일으켜, 경제적으로 막대한 손실을 발생한다. 특히, 병원성 미생물은 식중독, 말라리아, 톡소플라즈마병, 피부 사상균증, 피부 칸디다증, 아구창, 중이염, 성홍열, 부비강염, 세균성 폐렴, 세균성 장염, 전염성 농가진, 요로 감염증, 결핵, 코감기, 인후염, 돌방성 발진, 수두, 볼거리, 풍진, 수족구병, 전염성 홍반, 대상포진, 물사마귀 등과 같은 각종 질병을 유발시킨다.The harmful microorganisms distributed in the natural environment cause damage to the infection, resulting in a great economic loss. Particularly, the pathogenic microorganisms can be used for the treatment of diseases such as food poisoning, malaria, toxoplasmosis, dermatophytosis, cutaneous candidiasis, thrush, otitis, scarlet fever, sinusitis, bacterial pneumonia, bacterial enteritis, infectious pox, urinary tract infection, tuberculosis, Chickenpox, mumps, rubella, foot and mouth disease, infectious erythema, shingles, water warts and the like.
식중독은 "식품의 섭취로 인하여, 인체에 유해한 미생물 또는 유독물질에 의하여 발생하였거나, 발생한 것으로 판단되는 감염성 또는 독소형 질환"이라고 정의된다. 식중독을 일으키는 유해 세균으로는 스타필로코커스 아우레우스(Staphylococcus aureus), 바실러스 세레우스(Bacillus cereus), 리스테리아 모노사이토제니스(Listeria monocytogenes), 살모넬라 타이피(Salmonella typhi) 등이 있다. 이중, 바실러스 세레우스(Bacillus cereus)는 구토와 설사 모두 유발하는 맹독성 식중독균으로 알려져 있다. 식중독균은 여러 환경에 두루 분포되어 있어서, 식물을 원재료로 하는 음식이나, 고기류, 유제품, 쌀, 파스타 등에서 서식할 수 있다. Food poisoning is defined as "an infectious or poisonous disease caused by, or caused by, microorganisms or toxic substances harmful to the human body due to ingestion of food". Cause food poisoning by harmful bacteria is Staphylococcus aureus (Staphylococcus aureus), Bacillus cereus (Bacillus cereus), Listeria monocytogenes Zenith (Listeria monocytogenes ), Salmonella typhi typhi ). Among them, Bacillus cereus is known as a toxic food poisoning causing both vomiting and diarrhea. Foodborne pathogens are distributed throughout many environments and can be found in food made from plants, meat, dairy products, rice, and pasta.
미국에서는 연간 약 84,000건의 식중독이 바실러스 세레우스에 의해 발생하는 것으로 보고되고 있으며, 우리나라에서는 2002년부터 2010년 4월까지 약 27건의 집단식중독과 793명의 환자가 보고되어 있다. 하지만, 바실러스 세레우스의 감염으로 발생되는 실 환자수는 보고된 환자수와 비교하여 약 10~20배 이상 많이 발생되는 것으로 알려져 있다. In the United States, about 84,000 foodborne illnesses per year are reported to be caused by Bacillus cereus. In Korea, about 27 cases of group food poisoning and 793 cases were reported from 2002 to April 2010. However, it is known that the number of actual cases caused by infection with Bacillus cereus is about 10 to 20 times higher than the reported number of patients.
한편, 자연계에 존재하는 미생물은 인간에게 뿐만 아니라, 동물 및 식물에도 영향을 끼친다. 예를 들어 바실러스 세레우스(Bacillus cereus)는 토양, 수중, 공기 및 자연계에 널리 분포하고 있어, 대부분의 식물이 이에 영향을 받으며, 소, 돼지, 말, 토끼, 개와 같은 동물을 감염시키기도 한다. 또한, 리스테리아 모노사이토제니스(Listeria monocytogenes)의 경우, 동물을 감염시켜 신경계에 문제를 일으킬 수도 있다. On the other hand, microorganisms that exist in the natural world affect not only humans but also animals and plants. For example, Bacillus cereus is widely distributed in soil, water, air and the natural world, and most plants are affected by it and infect animals such as cattle, pigs, horses, rabbits and dogs. In addition, in the case of Listeria monocytogenes , it can infect animals and cause problems in the nervous system.
현재까지 화학적으로 합성한 항균제를 사용하여 식중독균을 포함한 여러 미생물을 제어하려는 시도는 많이 이루어져 왔지만, 합성 보존료를 식품에 직접 첨가하기 위해서는 위해 평가 과정을 거쳐서 인체에 독성이 없다는 사실이 증명되어야 한다. 또한, 일반적으로 소비자들은 합성보존료가 포함된 식품을 선호하지 않는 성향을 보이기 때문에 합성보존료는 사용에 많은 제한이 따른다. To date, many attempts have been made to control microorganisms including foodborne pathogens by using chemically synthesized antimicrobial agents. However, in order to add synthetic preservatives directly to foods, it has to be proved that they are not toxic to humans through a risk assessment process. In addition, synthetic preservatives have a lot of limitations because consumers tend to prefer foods that contain synthetic preservatives.
따라서, 사람 및 동식물에게 유해한 각종 미생물을 제어하기 위해서는 천연물 유래의 항균제를 개발하려는 노력이 필요한 실정이다.
Therefore, in order to control various microorganisms harmful to humans and plants and animals, efforts to develop antimicrobial agents derived from natural materials are needed.
본 발명은 여러 유해균에 대해 높은 생육 저해능을 나타내는 생육저해제를 개발하여 제공하는데 목적이 있다.
It is an object of the present invention to develop and provide a growth inhibitor exhibiting a high growth inhibitory effect against various noxious bacteria.
상기 목적을 달성하기 위하여, 본 발명은 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 또는 그 배양액을 제공한다. In order to achieve the above object, the present invention is Bacillus amyl Lowry query Pacific Enschede (Bacillus amyloliquefaciens s KNU-1 (KCTC18343P) or a culture thereof.
또한, 본 발명은 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 또는 그 배양액을 유효성분으로 함유하는 것을 특징으로 하는 미생물 생육저해제를 제공한다. 이때, 상기 미생물은 바람직하게 스타필로코커스 아우레우스(Staphylococcus aureus), 바실러스 세레우스(Bacillus cereus), 리스테리아 모노사이토제니스(Listeria monocytogenes), 스포로볼로마이세스 로제우스(Sporobolomyces roseus), 웨이셀라 비리데스켄스(Weissella viridescens), 스타필로코커스 에피데르미데스(Staphylococcus epidermidis) 및 살모넬라 타이피(Salmonella typhi) 중 선택되는 어느 하나일 수 있다.In addition, the present invention is Bacillus amyl Lowry query Pacific Enschede (Bacillus amyloliquefaciens s KNU-1 (KCTC18343P) or a culture thereof as an effective ingredient. At this time, the microorganisms are preferably Staphylococcus aureus (Staphylococcus aureus), Bacillus cereus (Bacillus cereus), Listeria monocytogenes Zenith (Listeria monocytogenes , Sporobolomyces roseus , Weissella viridescens , Staphylococcus epidermidis , and Salmonella typhi . The term " monocytogenes " .
본 발명의 상기 미생물 생육저해제는 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 또는 그 배양액을 유효성분으로 함유하고, 그 외에 약리 효능의 증진, 제제화 등을 위해 약제학적으로 필요한 부형제, 첨가제 등을 첨가할 수 있다. 이때, 약리효능의 증진 및 제제화를 위해 필요한 사항은 당업계의 통상적인 기술을 이용할 수 있으므로 이에 관한 기재는 생략하기로 한다. The microorganism growth inhibitor of the present invention contains Bacillus amyloliquefaciens KNU-1 (KCTC18343P) strain or a culture thereof as an active ingredient, and further contains pharmacologically necessary Excipients, additives, and the like may be added. At this time, the matters necessary for the promotion and formulation of the pharmacological efficacy can be obtained by conventional techniques in the art, so the description thereof will be omitted.
한편, 상기 스타필로코커스 아우레우스(Staphylococcus aureus)는 자연계에 널리 분포되어 있는 세균의 하나로서, 식중독 뿐 아니라 화농, 중이염, 방광염과 같은 화농성 질환을 일으키는 세균이다. On the other hand, Staphylococcus aureus is one of bacteria widely distributed in nature, and is a bacterium that causes not only food poisoning but also purulent diseases such as pneumonia, otitis media and cystitis.
또한, 상기 바실러스 세레우스(Bacillus cereus)는 토양, 먼지, 하수 등 자연계에 널리 분포하는 병원성 미생물의 일종으로 주로 식물에 오염되어 부패, 변패 및 식중독을 일으킬 수 있다. 바실러스 세레우스는 토양세균의 일종으로 자연계에 널리 분포하므로 농작물을 비롯한 대부분의 식품은 이 균에 의해 오염되어 있다고 볼 수도 있다. 따라서, 거의 모든 종류의 식품이 바실러스 세레우스 식중독과 연관이 있다고 할 수 있다. In addition, Bacillus cereus is a kind of pathogenic microorganisms widely distributed in the natural world such as soil, dust, sewage, etc., and is mainly contaminated with plants, and can cause decay, deterioration and food poisoning. Bacillus cereus is a kind of soil bacteria that is widely distributed in nature, so most foods, including crops, may be contaminated by this bacterium. Thus, almost all types of food are associated with Bacillus cereus food poisoning.
또한, 상기 리스테리아 모노사이토제니스(Listeria monocytogenes)는 수류 및 조류에 자연감염하는 균으로, 저온에서도 성장 가능하며, 사람에게 감염되어 리스테리아증을 유발시킬 수 있다. 리스테리아 모노사이토제니스에 오염된 음식을 섭취한 후, 12시간이 지나면 일반적인 식중독 증상(메스꺼움, 구토, 설사)인 위장관계 질환이 발생하고, 며칠 뒤에는 패혈증, 뇌수막염을 포함한 심각한 증세가 나타날 수 있다. Also, the zenith L. monocytogenes (Listeria monocytogenes) may be possible to grow bacteria in natural water stream and infect the birds, cold, infected and those who cause listeriosis. Gastrointestinal conditions such as common food poisoning symptoms (nausea, vomiting, diarrhea) occur within 12 hours after ingestion of food contaminated with Listeria monocytogenes, and severe symptoms including sepsis and meningitis may occur a few days later.
한편, 상기 스포로볼로마이세스 로제우스(Sporobolomyces roseus)는 효모균의 일종으로 식물에 유사황반병(pseudo greasy)을 유발하는 것으로 보고되어 있다. On the other hand, Sporobolomyces roseus is a kind of yeast and has been reported to cause pseudo greasy to plants.
한편, 상기 웨이셀라 비리데스켄스(Weissella viridescens)는 과산화수소를 생사하는 세균으로, 육류를 감염시키는 것으로 알려져 있다. On the other hand, the Weissella viridescens is a bacterium that produces hydrogen peroxide and is known to infect meat.
한편, 상기 스타필로코커스 에피데르미데스(Staphylococcus epidermidis)는 두부, 겨드랑이, 콧구멍, 양팔 및 다리에서 많이 서식하는 세균으로, 이에 감염되면 복막염 및 요로계 질환이 발생할 수 있다. On the other hand, Staphylococcus epidermidis is a bacterium which is found in a lot of tofu, armpits, nostrils, arms and legs. When infected, peritonitis and urinary tract diseases may occur.
한편, 상기 살모넬라 타이피(Salmonella typhi)는 주모성의 편모를 가지고 운동성을 나타내는 장내세균으로, 이에 감염되면 식중독, 장염, 고열, 두통, 불쾌감, 식욕감퇴가 발생할 수 있다. Meanwhile, the Salmonella typhi is a enterobacteria exhibiting motility with flagella of the main maternal, and when it is infected, food poisoning, enteritis, high fever, headache, discomfort, and loss of appetite may occur.
한편, 본 발명은 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 또는 그 배양액을 함유하는 식품첨가제 또는 식품을 제공한다. 상기 식품첨가제 또는 식품은 상기 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 또는 그 배양액를 함유하기만 하면 그 제형에 특별히 국한되지 않으나, 특별히 미생물에 오염되기 쉬운 식품에 더 잘 적용될 수 있다. On the other hand, the present invention is Bacillus amyl Lowry query Pacific Enschede (Bacillus Amyloliquefaciens s KNU-1 (KCTC18343P) or a food additive or food containing the culture. The food additive or food may be selected from the group consisting of Bacillus amyloliquefaciens amyloliquefaciens s KNU-1 (KCTC18343P) strain or a culture thereof is not particularly limited to the formulation, but may be more applicable to foods particularly susceptible to microbial contamination.
한편, 본 발명은 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 또는 그 배양액을 함유하는 사료첨가제 또는 사료를 제공한다. 상기 사료첨가제 또는 사료는 상기 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 또는 그 배양액을 함유하기만 하면 그 제형에 특별히 국한되지 않으나, 특별히 미생물에 오염되기 쉬운 사료에 더 잘 적용될 수 있다. 상기 사료첨가제 또는 사료 제조시, 부가적으로 필요한 사항은 미생물 함유 사료의 제조에 관한 당업계의 공지기술을 활용할 수 있다. On the other hand, the present invention is Bacillus amyl Lowry query Pacific Enschede (Bacillus amyloliquefaciens s KNU-1 (KCTC18343P) strain or a culture thereof. The feed additive or feed may be selected from the group consisting of Bacillus amyloliquefaciens amyloliquefaciens s KNU-1 (KCTC18343P) strain or a culture thereof is not particularly limited to the formulation, but can be applied more particularly to feeds susceptible to microbial contamination. In addition, when preparing the feed additive or the feed, additional necessary matters may be utilized in the production of microorganism-containing feeds.
한편, 본 발명은 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 또는 그 배양액을 함유하는 비료첨가제 또는 비료를 제공한다. 상기 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 또는 그 배양액을 함유하고 있는 비료첨가제 또는 비료는 작물 재배시 병원성 세균의 증식을 억제하여 감염으로부터 식물을 보호할 수 있다. 상기 비료 첨가제 또는 비료 제조시 부가적으로 필요한 사항은 미생물 함유 비료 제조에 관한 당업계의 공지기술을 활용할 수 있다.
On the other hand, the present invention is Bacillus amyl Lowry query Pacific Enschede (Bacillus amyloliquefaciens s KNU-1 (KCTC18343P) strain or a culture thereof. The above-mentioned amyloliquefaciens ( Bacillus amyloliquefacien s) Fertilizer additives or fertilizers containing KNU-1 (KCTC18343P) strain or its culture can protect plants against infection by inhibiting the growth of pathogenic bacteria during crop cultivation. Any additional requirements for the production of the fertilizer additive or the fertilizer may utilize a well-known technique in the art for manufacturing microorganism-containing fertilizer.
본 발명의 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주는 여러 유해균에 대해 높은 생육 저해능을 나타내기 때문에, 식품, 사료, 비료 등에 첨가되어 유해균의 생육을 억제하는 용도로 사용될 수 있다. Since the Bacillus amyloliquefaciens KNU-1 (KCTC18343P) strain of the present invention exhibits high growth inhibitory effect against various harmful bacteria, it is used for suppressing the growth of harmful bacteria added to food, feed, fertilizer and the like Can be used.
도 1은 본 발명의 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주의 유해균에 대한 생육저해능을 분석하고자 제조한 배지이다. 배지의 1번에는 본 발명의 'Bacillus amyloliquefaciens KNU-1 (KCTC18343P)', 2번에는 'Bacillus amyloliquefaciens KCTC1660', 3번에는 'Bacillus amyloliquefaciens KCTC2105', 4번에는 'Bacillus amyloliquefaciens KCTC3002', 5번에는 'Bacillus amyloliquefaciens KCTC13588'가 접종되어 있으며, NC로 표시된 부분은 음성대조군인 상태이다.
도 2는 '스타필로코커스 아우레우스 KCCM12214'에 대한 본 발명 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주의 생육억제능 결과이다.
도 3은 '스타필로코커스 아우레우스 KCTC1916'에 대한 본 발명 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주의 생육억제능 결과이다.
도 4는 '바실러스 세레우스 KCTC1012'에 대한 본 발명 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주의 생육억제능 결과이다.
도 5는 '리스테리아 모노사이토제니스 ATCC15313'에 대한 본 발명 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주의 생육억제능 결과이다.
도 6은 '스타필로코커스 아우레우스 KCCM12214'에 대한 본 발명 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 배양액의 생육억제능 결과이다.
도 7은 '스타필로코커스 아우레우스 KCTC1916'에 대한 본 발명 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 배양액의 생육억제능 결과이다.
도 8은 '바실러스 세레우스 KCTC1012'에 대한 본 발명 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 배양액의 생육억제능 결과이다.
도 9는 '리스테리아 모노사이토제니스 ATCC15313'에 대한 본 발명 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 배양액의 생육억제능 결과이다.
도 10은 '스포로볼로마이세스 로제우스 KCTC17112'에 대한 본 발명 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 배양액의 생육억제능 결과이다.
도 11은 '웨이셀라 비리데스켄스 KCTC3504'에 대한 본 발명 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 배양액의 생육억제능 결과이다.
도 12는 '스타필로코커스 에피데르미데스 KCTC1917'에 대한 본 발명 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 배양액의 생육억제능 결과이다.
도 13은 '살모넬라 타이피 ATCC19430'에 대한 본 발명 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 배양액의 생육억제능 결과이다. Brief Description of the Drawings Fig. 1 is a medium prepared for the analysis of the inhibitory effect of the Bacillus amyloliquefaciens KNU-1 (KCTC18343P) strain of the present invention on the growth of harmful bacteria. In No. 1 of the medium, ' Bacillus amyloliquefaciens s KNU-1 (KCTC18343P) ', 2' Bacillus amyloliquefaciens KCTC1660 ', 3' Bacillus amyloliquefacien s KCTC2105 ', 4 times, the "Bacillus amyloliquefacien s KCTC3002', 5 time, 'and the Bacillus amyloliquefacien s KCTC13588' is inoculated, the portion indicated by the NC is the negative control condition.
FIG. 2 is a graph showing the effect of the present invention Bacillus amyloliquefaciens ( Bacillus subtilis) against 'Staphylococcus aureus KCCM12214' amyloliquefacien s) KNU-1 (KCTC18343P).
Figure 3 is a "Staphylococcus aureus KCTC1916, the invention Bacillus amyl Lowry query Pacific Enschede (Bacillus for amyloliquefacien s) KNU-1 (KCTC18343P).
4 is "Bacillus cereus KCTC1012, the invention Bacillus amyl Lowry query Pacific Enschede (Bacillus for amyloliquefacien s) KNU-1 (KCTC18343P).
Figure 5 is a graph showing the effect of the present invention Bacillus amyloliquefaciens ( Bacillus amyloliquefaciens) against 'Listeria monocytogenes ATCC 15313' amyloliquefacien s) KNU-1 (KCTC18343P).
FIG. 6 is a graph showing the effect of the present invention Bacillus amyloliquefaciens ( Bacillus subtilis) against 'Staphylococcus aureus KCCM 12214' amyloliquefacien s) KNU-1 (KCTC18343P).
FIG. 7 is a graph showing the effect of the present invention Bacillus amyloliquefaciens ( Bacillus subtilis) against 'Staphylococcus aureus KCTC1916' amyloliquefacien s) KNU-1 (KCTC18343P).
8 is a "Bacillus cereus KCTC1012, the invention Bacillus amyl Lowry query Pacific Enschede (Bacillus for amyloliquefacien s) KNU-1 (KCTC18343P).
Figure 9 is a graph showing the effect of the present invention Bacillus amyloliquefaciens ( Bacillus amyloliquefaciens) against ' Listeria monocytogenes ATCC 15313 & amyloliquefacien s) KNU-1 (KCTC18343P).
FIG. 10 is a graph showing the activity of & KCTC17112 'the present invention on Bacillus amyl Lowry query Pacific Enschede (Bacillus amyloliquefacien s) KNU-1 (KCTC18343P).
Figure 11 is a graphical representation of & KCTC3504 'the present invention on Bacillus amyl Lowry query Pacific Enschede (Bacillus amyloliquefacien s) KNU-1 (KCTC18343P).
FIG. 12 is a graph showing the relationship between " Staphylococcus epidermidis KCTC1917 'the present invention on Bacillus amyl Lowry query Pacific Enschede (Bacillus amyloliquefacien s) KNU-1 (KCTC18343P).
13 is the present invention, the Bacillus amyl Lowry query Pacific Enschede (Bacillus for "Salmonella tie blood ATCC19430" amyloliquefacien s) KNU-1 (KCTC18343P).
이하, 본 발명의 내용을 하기 실시예를 통해 더욱 상세히 설명하고자 한다. 다만, 본 발명의 권리범위가 하기 실시예에만 한정되는 것은 아니고, 그와 등가의 기술적 사상의 변형까지를 포함한다.
Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the scope of the present invention is not limited to the following embodiments, and includes modifications of equivalent technical ideas.
[실시예 1: 바실러스 아밀로리쿼파시엔스([Example 1: Preparation of Bacillus amyloliquefaciens ( Bacillus amyloliquefacienBacillus amyloliquefacien s) KNU-1 (KCTC18343P) 균주 분리]s) KNU-1 (KCTC18343P) strain isolation]
병원성 균에 대해 우수한 생육 저해능을 가진 미생물을 선발하고자, 전국에서 44점의 발효식품을 수집하였으며, 그 중 막걸리로부터 균주를 분리하고자 하였다. 우선, 막걸리 100 ㎕를 멸균한 생리식염수 900 ㎕가 들어있는 1.5 ml 에펜도르프 튜브 (eppendorf tube)에 넣고, 10-6까지 단계 희석하였다. Forty - four fermented foods were collected from all over the country in order to isolate microorganisms with pathogenic microorganisms. First, 100 μl of makgeolli was placed in a 1.5 ml eppendorf tube containing 900 μl of sterilized physiological saline and stepwise diluted to 10 -6 .
각각의 희석된 생리식염수 100 ㎕를 TSA 배지(40 g/l Trypton soy agar, Difco)에 분주한 후, 도말봉을 이용하여 도말하였다. 도말한 배지를 30℃, 암조건에서 36시간 정치배양하였다. 콜로니가 형성되면 형태나 색 혹은 특성에 따라 육안상 구분하고, 새로운 TSA 배지에 획선배양하여 균을 분리하였다. 100 μl of each diluted physiological saline was dispensed in TSA medium (40 g / l Trypton soy agar, Difco), and then plated using a smear rod. The stained medium was incubated for 36 hours at 30 DEG C under dark conditions. When colonies were formed, they were visually distinguished according to their shape, color, or characteristic, and they were isolated by a new strain on TSA medium.
분리한 균 중 여러 유해균들에 대해 생육 저해능이 가장 우수하게 나타난 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) 균주를 선별하고, 이를 '바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNI-1'로 명명한 후, 한국생명공학연구원 미생물자원센터에 기탁하여, 기탁번호 KCTC18343P를 부여받았다. Bacillus amyloliquefaciens ( Bacillus amyloliquefaciens), which exhibits the best growth inhibitory effect against various harmful bacteria among the isolated bacteria, amyloliquefacien s) selecting the strain, and this "Bacillus amyl Lowry query Pacific Enschede (Bacillus amyloliquefaciens s) KNI-1 ', and deposited with the microorganism resource center of the Korea Research Institute of Bioscience and received the deposit number KCTC18343P.
한편, 분리된 균주는 15% 글리세롤이 첨가되어 있는 크라이요 튜브 (Cryo tube)에 넣어, -80℃에서 보관하였다.
On the other hand, the isolated strain was put into a cryo tube containing 15% glycerol and stored at -80 ° C.
[[ 실험예Experimental Example 1: One: 바실러스Bacillus 아밀로리쿼파시엔스Amylori quasi-life (( BacillusBacillus amyloliquefacienamyloliquefacien ss ) KNU-1 (KCTC18343P) 균주의 유해균에 대한 ) For KNU-1 (KCTC18343P) strain of harmful bacteria 생육저해능Growth inhibition 분석] analysis]
본 발명 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주의 유해균에 대한 생육저해능을 분석하고자 하였다. 도 1에서 보는 바와 같이, 각종 유해균이 도말된 배지 위, 1번 위치에는 본 발명의 '바실러스 아밀로리쿼파시엔스 KNU-1 (KCTC18343P)'를, 2번 위치에는 표준균주인 '바실러스 아밀로리쿼파시엔스 KCTC1660', 3번 위치에는 표준균주인 '바실러스 아밀로리쿼파시엔스 KCTC2105', 4번 위치에는 표준균주인 '바실러스 아밀로리쿼파시엔스 KCTC3002', 5번 위치에는 표준균주인 '바실러스 아밀로리쿼파시엔스 KCTC13588'을 접종한 후, 배양하여 유해균에 대한 생육저해 여부를 관찰하였다. NC로 표시된 부분은 음성대조군이다. The present invention Bacillus amyl Lowry query Pacific Enschede (Bacillus amyloliquefacien s) KNU-1 (KCTC18343P) against the pathogenic bacteria. As shown in FIG. 1, 'Bacillus amyloliquefaciens KNU-1 (KCTC18343P)' of the present invention was deposited at
먼저, 배지에 유해균으로 '스타필로코커스 아우레우스 KCCM12214', '스타필로코커스 아우레우스 KCTC1916', '바실러스 세레우스 KCTC1012', '리스테리아 모노사이토제니스 ATCC15313'의 4가지 균주를 각각 도말한 후, 도 1에서와 같은 위치에 상기 1~5번의 바실러스 아밀로리쿼파시엔스 균주를 접종한 후, 유해균에 대한 생육저해능을 관찰하였다. First, four strains of Staphylococcus aureus KCCM12214, Staphylococcus aureus KCTC1916, Bacillus cereus KCTC1012 and Listeria monocytogenes ATCC15313 were streaked into the culture medium, After inoculation of the
실험 결과, 도 2, 3, 4 및 5로부터 확인되는 바와 같이, 유해균 '스타필로코커스 아우레우스 KCCM12214', '스타필로코커스 아우레우스 KCTC1916' 및 '리스테리아 모노사이토제니스 ATCC15313'에 대해, 본 발명 바실러스 아밀로리쿼파시엔스 KNU-1 (KCTC18343P) 균주의 생육저해능이 다른 바실러스 아밀로리쿼파시엔스 균주와 대비하여 볼 때 가장 뚜렷하게 나타났다.
As can be seen from the results of FIGS. 2, 3, 4, and 5, the present inventors have found that the present invention can be applied to the pests of Staphylococcus aureus KCCM 12214, Staphylococcus aureus KCTC1916 and Listeria monocytosineus ATCC 15313, The growth inhibitory effect of Bacillus amyloliquefaciens KNU-1 (KCTC18343P) strain was most pronounced when compared to other Bacillus amyloliquefaciens strains.
[[ 실험예Experimental Example 2: 2: 바실러스Bacillus 아밀로리쿼파시엔스Amylori quasi-life (( BacillusBacillus amyloliquefacienamyloliquefacien ss ) ) KNUKNU -1 (KCTC18343P) 균주 배양액의 유해균에 대한 -1 (KCTC18343P) for the noxious bacteria of the culture broth 생육저해능Growth inhibition 분석] analysis]
본 실험예에서는 본 발명의 바실러스 아밀로리쿼파시엔스(Bacillus amyloliquefaciens) KNU-1 (KCTC18343P) 균주 배양액의 유해균에 대한 생육저해능을 분석하고자 하였다. 도 1과 같이 유해균이 도말된 배지 위, 1번에는 본 발명의 '바실러스 아밀로리쿼파시엔스 KNU-1 (KCTC18343P)'를, 2번에는 표준균주인 '바실러스 아밀로리쿼파시엔스 KCTC1660', 3번에는 표준균주인 '바실러스 아밀로리쿼파시엔스 KCTC2105', 4번에는 표준균주인 '바실러스 아밀로리쿼파시엔스 KCTC3002', 5번에는 표준균주인 '바실러스 아밀로리쿼파시엔스 KCTC13588'을 접종하였다. NC로 표시된 부분은 음성대조군이다. In this Example, the growth inhibitory effect of the culture medium of Bacillus amyloliquefaciens KNU-1 (KCTC18343P) of the present invention on the pathogenic bacteria was analyzed. Bacillus amyloliquefaciens KNU-1 (KCTC18343P) 'of the present invention was used as the first strain,' Bacillus amyloliquefaciens KCTC1660 ', 3 strains used as the second strain, Bacillus amyloliquefaciens KCTC2105 'as a standard strain,' Bacillus amyloliquefaciens KCTC3002 'as a standard strain in No. 4, and' Bacillus amyloliquefaciens KCTC13588 'as a standard strain in 5th. The area labeled NC is the negative control.
유해균으로는 '스타필로코커스 아우레우스 KCCM12214', '스타필로코커스 아우레우스 KCTC1916', '바실러스 세레우스 KCTC1012', '리스테리아 모노사이토제니스 ATCC15313', '스포로볼로마이세스 로제우스 KCTC17112', '웨이셀라 비리데스켄스 KCTC3504', '스타필로코커스 에피데르미데스 KCTC1917', '살모넬라 타이피 ATCC19430'의 8가지 균주를 사용하였다. Examples of the harmful bacteria include 'Staphylococcus aureus KCCM12214', 'Staphylococcus aureus KCTC1916', 'Bacillus cereus KCTC1012', 'Listeria monocytogenes ATCC15313', 'Sporobolomiases rozeus KCTC17112 "," Weisselavir desksen KCTC3504 ',' Staphylococcus epidermidis KCTC1917 " and " Salmonella typhi ATCC19430 ".
각각의 유해균을 배지에 도말한 후, 도 1에서와 같이 1~5번의 바실러스 아밀로리쿼파시엔스 균주를 접종한 후, 유해균을 배양하였다. Each of the noxious bacteria was spread on a medium and inoculated with 1 to 5 Bacillus amyloliquefaciens strains as shown in Fig. 1, and then the noxious bacteria were cultured.
실험결과, 도 6, 7, 8, 9, 10, 11, 12 및 13으로부터 확인되는 바와 같이 본 발명의 바실러스 아밀로리쿼파시엔스 KNU-1 (KCTC18343P) 균주는 '스타필로코커스 아우레우스 KCCM12214', '스타필로코커스 아우레우스 KCTC1916', '바실러스 세레우스 KCTC1012', '리스테리아 모노사이토제니스 ATCC15313', '스포로볼로마이세스 로제우스 KCTC17112', '웨이셀라 비리데스켄스 KCTC3504', '스타필로코커스 에피데르미데스 KCTC1917', '살모넬라 타이피 ATCC19430'균에 대해 모두 생육저해능을 나타내었다.
6, 7, 8, 9, 10, 11, 12 and 13, the Bacillus amyloliquefaciens KNU-1 (KCTC18343P) strain of the present invention was identified as 'Staphylococcus aureus KCCM12214' , 'Staphylococcus aureus KCTC1916', 'Bacillus cereus KCTC1012', 'Listeria monocytogenes ATCC 15313', 'Sporobolomiases rozeus KCTC17112 "," Weisselavir desksen KCTC3504 ',' Staphylococcus epidermidis KCTC1917 'and' Salmonella typhi ATCC19430 ', respectively.
[[ 제조예Manufacturing example 1 : One : 손세정제Hand cleaner 제조] Produce]
비이커에 에탄올 67.67 g, 향 0.05 g을 넣고, 증류수 24.33 g, 글리세린 0.5 g을 넣고 25℃에서 3,000 rpm 으로 1분간 교반시켰다. 여기에 프로필렌글리콜 0.5 g, 이소프로필알콜 1.5 g, 이소프로필미리스테이트 0.5 g, 올리고당 0.004 g, 키토산 0.004 g을 차례로 넣고 교반기로 1분 정도 교반하고, 교반하는 동안 디이소프로판올 아민 0.2 g을 가하여 교반하였다. 모든 성분이 완전히 녹은 것을 확인한 후, 색소 0.22 g 및 바실러스 아밀로리쿼파시엔스 KNU-1 (KCTC18343P) 균주의 배양액 (제균처리한 배양액) 10 ml을 첨가하고, 카보머 0.3 g을 넣고, 30분 정도 3,000rpm 으로 교반하여 젤을 제조하였다.
To the beaker, 67.67 g of ethanol and 0.05 g of fragrance were added, and 24.33 g of distilled water and 0.5 g of glycerin were added and stirred at 25 ° C for 3 minutes at 3,000 rpm for 1 minute. 0.5 g of propylene glycol, 1.5 g of isopropyl alcohol, 0.5 g of isopropyl myristate, 0.004 g of oligosaccharide and 0.004 g of chitosan were added in this order, and the mixture was stirred for about one minute with a stirrer. While stirring, 0.2 g of diisopropanolamine was added and stirred . After confirming that all the components were completely dissolved, 10 ml of a culture solution (culture solution with sterilization treatment) of 0.22 g of pigment and Bacillus amyloliquefaciens KNU-1 (KCTC18343P) was added, 0.3 g of carbomer was added, The mixture was stirred at 3,000 rpm to prepare a gel.
[[ 제조예Manufacturing example 2: 2: 바실러스Bacillus 아밀로리쿼파시엔스Amylori quasi-life KNU-1 (KCTC18343P) 균주의 배양액을 함유하는 식품첨가제의 제조] Preparation of Food Additives Containing Cultures of KNU-1 (KCTC18343P)
표고버섯 분말 50 g, 새우 분말 30 g, 다시마 분말 15 g 및 본 발명의 바실러스 아밀로리쿼파시엔스 KNU-1 (KCTC18343P) 균주의 배양액 (제균처리한 배양액) 5 ml를 혼합하여 조미료로 사용될 수 있는 식품첨가제를 제조하였다.
5 ml of a culture solution (culture solution obtained by sterilization treatment) of 50 g of mushroom powder, 30 g of shrimp powder, 15 g of kelp powder and the Bacillus amyloliquefaciens KNU-1 (KCTC18343P) strain of the present invention was mixed with the mixture Food additives were prepared.
[[ 제조예Manufacturing example 3: 3: 바실러스Bacillus 아밀로리쿼파시엔스Amylori quasi-life KNU-1 (KCTC18343P) 균주의 배양액을 함유하는 식품의 제조] Preparation of Food Containing Culture Medium of KNU-1 (KCTC18343P)
<우유 제조><Milk production>
시판 중인 S 제조의 우유 200 ml에 본 발명의 바실러스 아밀로리쿼파시엔스 KNU-1 (KCTC18343P) 균주의 배양액 (제균처리한 배양액) 2 ml를 첨가하여 우유 조성물을 제조하였다.A milk composition was prepared by adding 2 ml of a culture solution (culture solution obtained by sterilization) of the Bacillus amyloliquefaciens KNU-1 (KCTC18343P) strain of the present invention to 200 ml of commercially available S milk.
<바나나 쥬스 제조> <Banana juice manufacturing>
바나나를 믹서에 넣고 갈아 바나나 원액 180 ml를 준비한 후, 본 발명의 바실러스 아밀로리쿼파시엔스 KNU-1 (KCTC18343P) 균주의 배양액 (제균처리한 배양액) 2 ml를 첨가하여 바나나 쥬스 조성물을 제조하였다.
A banana juice composition was prepared by adding 2 ml of a culture solution (a culture solution obtained by sterilization treatment) of the Bacillus amyloliquefaciens KNU-1 (KCTC18343P) strain of the present invention after preparing 180 ml of a banana stock solution prepared by putting the banana into a mixer.
[ [ 제조예Manufacturing example 4: 4: 바실러스Bacillus 아밀로리쿼파시엔스Amylori quasi-life KNU-1 (KCTC18343P) 균주의 배양액을 함유하는 사료첨가제 제조] Preparation of Feed Additive Containing Culture Medium of KNU-1 (KCTC18343P)
α-토코페롤 5 g, 오징어간유 0.75 g, 효모 93.25 g, 본 발명의 바실러스 아밀로리쿼파시엔스 KNU-1 (KCTC18343P) 균주의 배양액 (제균처리한 배양액) 1 ml를 혼합하여 사료첨가제를 제조하였다.
a feed additive was prepared by mixing 5 g of alpha -tocopherol, 0.75 g of squid liver oil, 93.25 g of yeast, and 1 ml of a culture solution (a culture solution obtained by sterilization treatment) of the strain of Bacillus amyloliquefaciens KNU-1 (KCTC18343P) of the present invention.
[[ 제조예Manufacturing example 5: 5: 바실러스Bacillus 아밀로리쿼파시엔스Amylori quasi-life KNU-1 (KCTC18343P) 균주의 배양액을 함유하는 사료의 제조] Preparation of feed containing culture medium of KNU-1 (KCTC18343P) strain]
< 육우 사료조성물 제조><Preparation of beef feed composition>
기본사료는 미가공옥수수 5 kg, 면실피 펠렛 1.5 kg, 생미강 0.5 kg, 소맥피1.5 kg, 면실박 0.45 kg, 대두박 1.0 kg, 소금 0.1 kg, 석회석 0.15 kg, 당밀 0.25 kg, 미량광물질 0.05 kg으로 조제되었다. 여기에 본 발명의 바실러스 아밀로리쿼파시엔스 KNU-1 (KCTC18343P) 균주의 배양액 (제균처리한 배양액) 10 ml를 첨가하였다.The basic diet was 5 kg of raw corn, 1.5 kg of cottonseed pellets, 0.5 kg of raw fish, 1.5 kg of wheat bran, 0.45 kg of cottonseed meal, 1.0 kg of soybean meal, 0.1 kg of salt, 0.15 kg of limestone, 0.25 kg of molasses and 0.05 kg of trace minerals It was prepared. To this was added 10 ml of a culture solution (culture solution obtained by sterilization treatment) of the Bacillus amyloliquefaciens KNU-1 (KCTC18343P) strain of the present invention.
<산란계용 사료조성물 제조>≪ Preparation of feeding composition for laying hens &
본 발명의 바실러스 아밀로리쿼파시엔스 KNU-1 (KCTC18343P) 균주의 배양액 (제균처리한 배양액) 10 ml를 시판 산란계용 사료(디럭스 산란(제품명), 부국사료, 한국)에 첨가하여 산란계용 사료조성물 1 kg을 제조하였다.10 ml of the culture (the culture solution obtained by sterilization treatment) of the strain of Bacillus amyloliquefaciens KNU-1 (KCTC18343P) of the present invention was added to a feed for commercial laying hens (deluxe scattering (product name) 1 kg.
<넙치 양식용 사료조성물 제조><Preparation of feed composition for flounder cultivation>
어분 5.8 kg, 대두박 0.4 g, 콘글루텐분 0.3 g, 소맥분 2.87 g, 오징어간유 0.4 g, 비타민 혼합제 0.12 g 및 미네랄 혼합제 0.1 g을 포함한 기본 사료에, 본 발명의 바실러스 아밀로리쿼파시엔스 KNU-1 (KCTC18343P) 균주의 배양액 (제균처리한 배양액) 1 ml를 첨가하여 넙치 양식용 사료조성물을 제조하였다.
Bacillus amyloliquefaciens KNU-1 of the present invention was added to a basic feed containing 5.8 kg of fish meal, 0.4 g of soybean meal, 0.3 g of corn gluten meal, 2.87 g of wheat flour, 0.4 g of squid liver oil, 0.12 g of a vitamin mixture and 0.1 g of a mineral mixture. (KCTC18343P) culture broth (sterilized broth) was added to prepare a feed composition for flounder culture.
[제조예 6: 바실러스 아밀로리쿼파시엔스 KNU-1 (KCTC18343P) 균주를 함유하는 비료첨가제의 제조][Preparation Example 6: preparation of fertilizer additive containing Bacillus amyloliquefaciens KNU-1 (KCTC18343P) strain]
쇠비름 50 g, 들깨 20 g, 봉선화 15 g을 혼합한 후, 무산소 상태에서 300℃로 60분 가열하고, 35℃로 45분간 가공하는 탄화방법으로 불포화 지방산을 추출하고, 본 발명의 바실러스 아밀로리쿼파시엔스 KNU-1 (KCTC18343P) 균주 0.1 g을 첨가하여 비료첨가제를 제조하였다.
The unsaturated fatty acid was extracted by a carbonization method in which 50 g of Peppermint, 20 g of perilla and 15 g of Balsinacea were mixed and heated in an oxygen-free state at 300 ° C. for 60 minutes and at 35 ° C. for 45 minutes. 0.1 g of Fasciens KNU-1 (KCTC18343P) strain was added to prepare a fertilizer additive.
[[ 제조예Manufacturing example 7: 7: 바실러스Bacillus 아밀로리쿼파시엔스Amylori quasi-life KNU-1 (KCTC18343P) 균주를 함유하는 비료의 제조] Preparation of fertilizer containing KNU-1 (KCTC18343P) strain]
음식물쓰레기, 도축쓰레기, 어패류쓰레기를 혼합 파쇄하고 이물질을 제거하여 혼합 폐기물을 제조하고, 혼합 폐기물의 수분함량을 55중량%로 조정하였다. 그 후, 수분함량이 조정된 혼합 폐기물 50중량%에 복합비료 15중량%, 요소비료 10중량%, 황토 5중량%, 생석회 20중량%를 넣어 처리조 뚜껑을 봉쇄하고, 2시간 발열 반응시켰다. 발열 반응 후, 인출하여 안정화 및 파쇄하고, 본 발명의 바실러스 아밀로리쿼파시엔스 KNU-1 (KCTC18343P) 균주 1.0중량%를 첨가하여 비료를 제조하였다.
Food wastes, slaughtered garbage and fish and shell garbage were mixed and crushed, foreign matter was removed to prepare mixed waste, and the moisture content of the mixed waste was adjusted to 55% by weight. Thereafter, 15% by weight of the compound fertilizer, 10% by weight of urea fertilizer, 5% by weight of loess, and 20% by weight of burnt lime were added to 50% by weight of the mixed waste whose moisture content was adjusted, and the treatment tank lid was sealed and subjected to exothermic reaction for 2 hours. After the exothermic reaction, it was taken out, stabilized and crushed, and 1.0% by weight of Bacillus amyloliquefaciens KNU-1 (KCTC18343P) strain of the present invention was added to produce a fertilizer.
Claims (6)
For example, Staphylococcus aureus , Bacillus cereus , Listeria monocytogenes , Sporobolomyces roseus , Weissella viridescens , Bacillus amyloliquefaciens KNU-1 (KCTC18343P) strain or a culture thereof, characterized by having a growth inhibitory effect against Staphylococcus epidermidis and Salmonella typhi .
For example, Staphylococcus aureus , Bacillus cereus , Listeria monocytogenes , Sporobolomyces roseus , Weissella viridescens , Bacillus amyloliquefaciens KNU-1 (KCTC18343P) strain or a culture thereof, which is characterized by having a growth inhibitory ability against Staphylococcus epidermidis and Salmonella typhi , A microbial growth inhibitor characterized by containing as an active ingredient.
A food comprising the strain of Bacillus amyloliquefaciens s KNU-1 (KCTC18343P) of claim 1 or a culture thereof.
A feed comprising the Bacillus amyloliquefaciens s KNU-1 (KCTC18343P) strain of claim 1 or a culture thereof.
A fertilizer containing the Bacillus amyloliquefaciens KNU-1 (KCTC18343P) strain of claim 1 or a culture thereof.
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| KR20220063345A (en) * | 2020-11-10 | 2022-05-17 | 박정옥 | Method of manufacturing feed additives mixed with complex microorganism |
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