KR20190067527A - A Bacillus probiotics mixture for preventing or treating obesity, diabetes and fatty liver and uses thereof - Google Patents
A Bacillus probiotics mixture for preventing or treating obesity, diabetes and fatty liver and uses thereof Download PDFInfo
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- KR20190067527A KR20190067527A KR1020170167552A KR20170167552A KR20190067527A KR 20190067527 A KR20190067527 A KR 20190067527A KR 1020170167552 A KR1020170167552 A KR 1020170167552A KR 20170167552 A KR20170167552 A KR 20170167552A KR 20190067527 A KR20190067527 A KR 20190067527A
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Abstract
Description
본 발명은 비만, 당뇨와 지방간의 예방과 치료 효과를 갖는 바실러스 혼합 조성물 및 이의 용도에 관한 것으로, 보다 상세하게 비만, 당뇨와 지방간의 예방과 치료 효과를 갖는 바실러스 혼합 조성물, 이를 포함하는 프로바이오틱스 제제, 건강기능식품 조성물 및 약학적 조성물에 관한 것이다.The present invention relates to a Bacillus mixed composition having a preventive and therapeutic effect on obesity, diabetes and fatty liver, and its use, and more particularly to a Bacillus mixed composition having a preventive and therapeutic effect on obesity, diabetes and fatty liver, a probiotic preparation, Health functional food compositions and pharmaceutical compositions.
비만은 대부분의 국가에서 주요한 보건 상의 문제가 되고 있다. 세계보건 기구(WHO)는 전 세계적으로 10억 명 이상의 성인은 과체중이고, 이들 중 3억 명은 임상적으로 비만인 것으로 추정하고 있다(Hotamisligil, 2006). 이러한 비만은 심혈관 질환, 뇌졸중, 인슐린 내성, 당뇨병, 간 질환, 신경퇴행성 질환, 호흡기 질환 및 기타 중증 질환 등에 대한 주요한 위험 인자(Risk factor)이고, 유방암 및 결장암을 포함한 일부 암에 대한 위험인자인 것으로 확인되었다. 또한, 신체 건강에 대한 영향 이외에, 비만은 우울증, 식이장애 및 낮은 자긍심을 초래하여 삶의 질 및 정신 건강에 유해한 영향을 미친다.Obesity is a major health problem in most countries. The World Health Organization (WHO) estimates that over one billion adults worldwide are overweight, and 300 million of them are clinically obese (Hotamisligil, 2006). Such obesity is a major risk factor for cardiovascular disease, stroke, insulin resistance, diabetes, liver disease, neurodegenerative diseases, respiratory diseases and other serious diseases, and is a risk factor for some cancers including breast and colon cancer . In addition, in addition to physical health effects, obesity causes depression, dietary habits, and low self esteem, which have a detrimental effect on quality of life and mental health.
이러한 가운데, 높은 에너지 부하를 갖는 식품 섭취의 꾸준한 증가 및 적은 운동과 연관된 사회적 변화들은 비만 및 비만으로 야기된 대사성 질환의 발생을 증가시키고 있다. 그러나, 저칼로리 다이어트와 운동에 기초한 전통적인 치료는 비만 제어에 있어서 그다지 효과를 나타내지 못하고, 일시적인 체중 손실을 초래할 뿐이다.Among these, the steady increase in food intake with high energy loads and the social changes associated with less exercise increase the incidence of obesity and obesity-induced metabolic diseases. However, conventional treatments based on low-calorie diets and exercises have little effect on obesity control and result in temporary weight loss.
따라서, 비만의 예방과 치료 뿐만 아니라 비만으로 야기되는 대사성 질환의 예방과 치료 효과를 갖는 약물의 개발이 절실한 실정이다.Therefore, there is a need for prevention and treatment of obesity, as well as development of a drug having the effect of preventing and treating metabolic diseases caused by obesity.
본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 비만, 당뇨와 지방간의 예방과 치료 효과를 가짐으로써, 비만, 당뇨 그리고 이로 인해 야기되는 대사성 질환의 발생을 억제하는데 사용될 수 있는 바실러스 혼합 조성물을 제공한다.The present invention provides a bacillus mixed composition which can be used for inhibiting the occurrence of obesity, diabetes and metabolic diseases caused by obesity, diabetes and fatty liver, do.
여기서, 바실러스 혼합 조성물은 바실러스속 균주 5종을 포함하고, 된장으로부터 분리될 수 있다.Here, the Bacillus mixed composition contains five species of Bacillus sp. And can be isolated from doenjang.
본 발명의 일 실시예에 따른 바실러스 혼합 조성물은 비만 억제 효능을 가지며, 바실러스 소노렌시스(Bacillus sonorensis) JJY12-3 균주(기탁번호: KCTC13405BP), 바실러스 파라리체니포르미스(Bacillus paralicheniformis) JJY12-8 균주(기탁번호: KCTC13406BP), 바실러스 소노렌시스(Bacillus sonorensis) JJY13-1 균주(기탁번호: KCTC13407BP), 바실러스 소노렌시스(Bacillus sonorensis) JJY13-3 균주(기탁번호: KCTC13408BP), 바실러스 소노렌시스(Bacillus sonorensis) JJY13-8 균주(기탁번호: KCTC13409BP)를 포함한다.The Bacillus mixed composition according to an embodiment of the present invention has an obesity-suppressing effect, and is useful as an antioxidant for Bacillus sonorensis JJY12-3 (accession number: KCTC13405BP), Bacillus paralicheniformis JJY12-8 (Accession No .: KCTC13406BP), Bacillus sonorensis JJY13-1 (Accession No .: KCTC13407BP), Bacillus sonorensis JJY13-3 (Accession No .: KCTC13408BP), Bacillus sonorensis ( Bacillus sonorensis ) strain JJY13-8 (Accession No .: KCTC13409BP).
상기 바실러스 혼합 조성물은 당뇨 예방 효능 및 지방간 축적 억제 효능을 갖는 것을 특징으로 한다.The Bacillus mixed composition is characterized by having a diabetes-preventing effect and an anti-fat accumulation effect.
본 발명의 실시예는, 상기 바실러스 혼합 조성물, 상기 바실러스 혼합 조성물의 배양물, 상기 배양물의 농축액 및 건조물로 이루어지는 군에서 선택되는 하나 이상을 유효성분으로 포함하는 프로바이오틱스 제제를 제공한다.An embodiment of the present invention provides a probiotic preparation comprising, as an active ingredient, at least one selected from the group consisting of the Bacillus mixed composition, the culture of the Bacillus mixed composition, the concentrate of the culture, and the dried product.
본 발명의 실시예는, 상기 바실러스 혼합 조성물, 상기 바실러스 혼합 조성물의 배양물, 상기 배양물의 농축액 및 건조물로 이루어지는 군에서 선택되는 하나 이상을 유효성분으로 포함하는 건강기능식품 조성물을 제공한다.An embodiment of the present invention provides a health functional food composition comprising, as an active ingredient, at least one selected from the group consisting of the Bacillus mixed composition, the culture of the Bacillus mixed composition, the concentrate of the culture, and the dried product.
본 발명의 실시예는, 상기 바실러스 혼합 조성물, 상기 바실러스 혼합 조성물의 배양물, 상기 배양물의 농축액 및 건조물로 이루어지는 군에서 선택되는 하나 이상을 유효성분으로 포함하는 약학적 조성물을 제공한다.An embodiment of the present invention provides a pharmaceutical composition comprising at least one selected from the group consisting of the Bacillus mixed composition, the culture of the Bacillus mixed composition, the concentrate of the culture, and the dried material as an active ingredient.
본 발명의 실시예는, 상기 바실러스 혼합 조성물, 상기 바실러스 혼합 조성물의 배양물, 상기 배양물의 농축액 및 건조물 중 적어도 하나를 유효 성분으로 포함하는 음료를 제공한다.An embodiment of the present invention provides a beverage comprising at least one of the Bacillus mixed composition, the culture of the Bacillus mixed composition, the concentrate of the culture and the dried product as an active ingredient.
본 발명의 실시예는, 상기 바실러스 혼합 조성물, 상기 바실러스 혼합 조성물의 배양물, 상기 배양물의 농축액 및 건조물 중 적어도 하나를 유효 성분으로 포함하는 건강 보조 식품을 제공한다.An embodiment of the present invention provides a health supplement containing, as an active ingredient, at least one of the Bacillus mixed composition, the culture of the Bacillus mixed composition, the concentrate of the culture, and the dried product.
본 발명의 실시예들에 따르면, 바실러스 혼합 조성물은 비만, 당뇨와 지방간의 예방과 치료 효과를 가짐으로써, 비만, 당뇨 그리고 이로 인해 야기되는 대사성 질환의 발생을 억제하는데 사용될 수 있다.According to the embodiments of the present invention, the Bacillus mixed composition can be used for suppressing the occurrence of obesity, diabetes and metabolic diseases caused thereby by having the preventive and therapeutic effect of obesity, diabetes and fatty liver.
본 발명이 실시예에 따르면, 본 발명의 바실러스 혼합 조성물을 함유하는 기능성 식품 또는 약제학적 조성물은 몸무게 증가 및 간 지방 축적에 대한 억제효능을 나타내고 내당능 증대에 뛰어난 효능을 나타내기 때문에 기능성 식품 또는 의약 분야에서 매우 유용할 수 있다.According to the present invention, the functional food or pharmaceutical composition containing the Bacillus mixed composition of the present invention exhibits an inhibitory effect on body weight gain and hepatic fat accumulation and exhibits an excellent effect on the increase in the intestine glucose tolerance, Can be very useful.
도 1은 본 발명에서 분리된 바실러스속 균주에 대한 안전성 검사 결과를 나타낸 도면이다.
도 2는 본 발명에서 분리된 바실러스속 균주 5종의 당 발효능 평가 결과를 나타낸 도면이다.
도 3은 본 발명의 바실러스 혼합 조성물 섭취에 의한 체중 감소 효과를 나타낸 도면이다.
도 4는 본 발명의 바실러스 혼합 조성물 섭취에 의한 지방조직의 무게 감소 효과를 나타낸 도면이다.
도 5는 본 발명의 바실러스 혼합 조성물 섭취에 의한 내당능 증대 효과를 나타낸 도면이다.
도 6은 본 발명의 바실러스 혼합 조성물 섭취에 의한 혈중 인슐린 감소 효과를 나타낸 도면이다.
도 7은 본 발명의 바실러스 혼합 조성물 섭취에 의한 지방간 개선 효과를 나타낸 도면이다.Brief Description of the Drawings Fig. 1 is a view showing the results of safety tests on strains of Bacillus sp. Isolated in the present invention. Fig.
FIG. 2 is a graph showing the results of evaluating the activity of five kinds of Bacillus sp. Isolates isolated in the present invention.
FIG. 3 is a graph showing the effect of the present invention on body weight reduction by ingesting a Bacillus mixed composition. FIG.
FIG. 4 is a graph showing the effect of weight reduction of adipose tissue by ingesting the Bacillus mixed composition of the present invention. FIG.
FIG. 5 is a graph showing an effect of increasing the glucose tolerance by ingesting the Bacillus mixed composition of the present invention. FIG.
FIG. 6 is a graph showing the effect of the present invention on blood insulin reduction by ingesting a Bacillus mixed composition. FIG.
FIG. 7 is a graph showing the effect of the present invention on the improvement of fatty liver by ingesting a Bacillus mixed composition. FIG.
이하, 본 발명에 따른 실시예들을 첨부된 도면을 참조하여 상세하게 설명한다. 그러나 본 발명이 실시예들에 의해 제한되거나 한정되는 것은 아니며, 당업자는 이러한 실시예들을 용이하게 변경할 수 있다. 또한, 각 도면에 제시된 동일한 참조 부호는 동일한 부재를 나타낸다.Hereinafter, embodiments according to the present invention will be described in detail with reference to the accompanying drawings. However, the present invention is not limited or limited by the embodiments, and those skilled in the art can easily modify these embodiments. In addition, the same reference numerals shown in the drawings denote the same members.
또한, 본 명세서에서 사용되는 용어(terminology)들은 본 발명의 바람직한 실시예를 적절히 표현하기 위해 사용된 용어들로써, 이는 시청자, 운용자의 의도 또는 본 발명이 속하는 분야의 관례 등에 따라 달라질 수 있다. 따라서, 본 용어들에 대한 정의는 본 명세서 전반에 걸친 내용을 토대로 내려져야 할 것이다.In addition, terminology used herein is a term used for appropriately expressing the preferred embodiment of the present invention, which may vary depending on the audience, the intention of the operator, or the practice of the field to which the present invention belongs. Therefore, the definitions of these terms should be based on the contents throughout this specification.
본 발명의 바실러스 혼합 조성물에 포함되는 바실러스속 균주에 대한 분리 과정에 대해 설명하면, 포항시 상사리에서 제조한 전통된장에서 바실러스속 균주를 분리하는 과정으로, 12년도에 제조하여 4년간 발효시킨 전통된장, 13년도에 제조하여 3년간 발효시킨 전통된장을 각각 1g 씩 사용하여 108,107,106,105로 희석 후, 각 플레이트에 희석액 100μL를 LB 한천배지(BD Difco)에 도말하여 37도에 18시간 배양한다. 후보 균주는 육안으로 확인하여 서로 다른 콜로니를 선택하고, 해당 후보 균주가 단일 균주임을 확인하기 위하여 3번 스트리킹(streaking)을 반복하여 동일하게 LB 한천배지에 도말하고 37도에 18시간 배양한다. 각 균주가 단일 균주임을 확인한 후, 16S 염기서열을 기준으로 분리 동정하고, 마지막으로 같은 종 내 아종(strain)을 구별하기 위하여 API50CH/B kit를 사용하여 당 발효능을 평가하여, 본 발명의 혼합 조성물에 사용되는 균주가 서로 다른 종 및 아종임을 확인한다.The separation process for the Bacillus sp. Strain contained in the Bacillus mixed composition of the present invention is as follows. The process for isolating the Bacillus sp. Isolate from the traditional miso produced in Pohang City Sangsari is as follows. 10 8 , 10 7 , 10 6 , and 10 5 using 1 g each of conventional miso fermented for 13 years and fermented for 3 years, and 100 μL of the diluted solution was plated on LB agar medium (BD Difco) For 18 hours. Candidate strains were selected by visual inspection and different colonies were selected. Streaking was repeated 3 times to confirm that the candidate strains were single strains, and the same strains were plated on LB agar medium and incubated at 37 ° C for 18 hours. After confirming that each strain was a single strain, the isolates were identified based on the 16S nucleotide sequence. Finally, the efficacy of the API50CH / B kit was evaluated to distinguish the subspecies of the same species, It is confirmed that the strains used in the composition are different species and subspecies.
여기서, 전통된장에서 분리된 바실러스속 균주는 17종이며, 이에 대한 것은 도 1에서 참조하여 설명한다.Here, there are 17 strains of Bacillus spp. Isolated from traditional miso, which will be described with reference to Fig.
바실러스 균주의 가장 대표적인 잠재적 독성 인자는 용혈 반응과 레시틴 분해 효소 발현이다. 해당 독성 인자 보유 여부를 확인하기 위하여, 포항지역 전통된장에서 분리한 바실러스속 균주 17종에 대하여 미생물 용혈성 검사(Hemolysis test)를 실시하고, 용혈성 검사 결과 음성으로 나타난 균주를 대상으로 레시틴아제 외독소 생성 검사(Lecithinase test)를 실시하여 각 균주의 안전성을 확인한다.The most prominent potential toxic factors of Bacillus strains are hemolytic reaction and lecithin degrading enzyme expression. In order to confirm the presence of the toxic factor, microorganism hemolysis test was performed on 17 strains of Bacillus spp. Isolated from traditional Doenjang in Pohang, and the strains negative for hemolytic test were examined for production of lecithinase exotoxin (Lecithinase test) to confirm the safety of each strain.
전통된장에서 분리된 바실러스속 균주에 대한 실험에 대해 간략하게 설명하면 다음과 같다.Experiments on strains of Bacillus sp. Isolated from traditional doenjang will be briefly described as follows.
미생물 microbe 용혈성검사Hemolytic test
TSA(tryptic soy agar) 배지에 5% 면양 혈액을 첨가하여 제조한 혈액한천배지에 16시간 이상씩, 2회 이상 37도로 호기 계대배양된 종균 후보균주를 도말하여 β hemolysis(완전용혈)와 α hemolysis(불완전용혈)를 측정한다.Bacterial candidate strains were cultured for more than 16 hours at 37 ° C for more than 16 hours in TSA (tryptic soy agar) medium supplemented with 5% fetal bovine serum. Β hemolysis (complete hemolysis) and α hemolysis (Incomplete hemolysis) is measured.
LecithinaseLecithinase 검사( inspection( 레시틴아제Lecithinase 외독소 생성 검사) Exotoxin production test)
MYP(Mannitol Egg Yolk Polymyxin) 한천배지를 46g/900ml의 농도로 희석하여 멸균한 후, Egg Yolk Enrichment(50%) 50 ml과 polymyxin B 4.1 ml(25,000 unit)를 첨가하여 제조한 배지에 종균 후보균주를 도말, 배지가 분홍색으로 변하고 clear zone을 만들면 Lecithinase 양성으로 판정하고, 노란색으로 변하면 Mannitol 양성으로 판정한다.MYP (Mannitol Egg Yolk Polymyxin) agar medium was diluted to a concentration of 46 g / 900 ml and sterilized. To the medium prepared by adding 50 ml of Egg Yolk Enrichment (50%) and 4.1 ml (25,000 units) of polymyxin B, Lecithinase positive is determined when the medium changes to pink, clear zone is made, and Mannitol is judged positive if it turns yellow.
도 1은 본 발명에서 분리된 바실러스속 균주에 대한 안전성 검사 결과를 나타낸 도면으로, 전통된장에서 분리된 바실러스속 균주 17종에 대한 안전성 검사 결과를 나타낸 도면이다.Brief Description of the Drawings Fig. 1 is a diagram showing the results of safety tests on strains of Bacillus sp. Isolated in the present invention. Fig. 1 shows the results of safety tests on 17 strains of Bacillus sp.
도 1에 도시된 바와 같이, 분리된 바실러스속 균주 17종 중 용혈성 검사와 레시틴아제 외독소 생성 검사를 통과한 균주는 바실러스 소노렌시스(Bacillus sonorensis) JJY12-3, 바실러스 파라리체니포르미스(Bacillus paralicheniformis) JJY12-8, 바실러스 소노렌시스(Bacillus sonorensis) JJY13-1, 바실러스 소노렌시스(Bacillus sonorensis) JJY13-3, 바실러스 소노렌시스(Bacillus sonorensis) JJY13-8인 것을 알 수 있다.As shown in FIG. 1, among the 17 strains of the Bacillus sp . Isolates, the strains that passed the hemolytic test and the test for the production of lecithinase exotoxin were Bacillus sonorensis JJY12-3, Bacillus paralicheniformis ) JJY12-8, Bacillus sonorensis JJY13-1, Bacillus sonorensis JJY13-3, and Bacillus sonorensis JJY13-8.
이렇게 안전성 검사를 통과한 바실러스 소노렌시스(Bacillus sonorensis) JJY12-3, 바실러스 파라리체니포르미스(Bacillus paralicheniformis) JJY12-8, 바실러스 소노렌시스(Bacillus sonorensis) JJY13-1, 바실러스 소노렌시스(Bacillus sonorensis) JJY13-3, 바실러스 소노렌시스(Bacillus sonorensis) JJY13-8의 5 균주를 1:1:1:1:1(1 = 1X108 CFU)로 혼합하여 바실러스 혼합 조성물을 형성한다. Bacillus sonorensis JJY12-3, Bacillus paralicheniformis JJY12-8, Bacillus sonorensis JJY13-1, Bacillus sonolensis, Bacillus subtilis, sonorensis) JJY13-3, Bacillus Sono alkylene sheath (Bacillus sonorensis) 5 strain of JJY13-8 1: 1: 1: 1 : 1 (1 = 1X10 8 CFU were mixed in a) to form a mixed composition of Bacillus.
여기서, CFU(colony forming unit)는 세균수 측정 단위를 의미한다.Here, CFU (colony forming unit) means a unit for measuring the number of bacteria.
바실러스 혼합 조성물을 구성하는 바실러스속 균주 5종 16S 염기서열은 다음과 같다.The 16S base sequences of the 5 species of Bacillus sp. Strain constituting the Bacillus mixed composition are as follows.
Bacillus Bacillus sonorensissonorensis JJY12JJY12 -3 -3
GGGTCCGATGTCGGATATTGGGCGTAAGCGCGCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGGAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAGAGGGTTTCCGCCCTTTAGTGCTGCAGCAAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTCTGACACCCCTAGAGATAGGGCTTCCCCTTCGGGGGCAGAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGCAGAACAAAGGGCAGCGAAGCCGCGAGGCTAAGCCAATCCCACAAATCTGCTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTGGAGCCAGCCGCCGAAGTGGGACAGATGATTGGGGTGAGCTAAAAGGAGTAGCCACAGCTAGTAAGGAGGTAGGCTTTCGCGCTCAGCGTCAGTTACAGACCAGAGAGTCGCCTTCGCCACTGGTGTTCCTCCACATCTCTACGCATTTCACCGCTACACGTGGAATTCCACTCTCCTCTTCTGCACTCAAGTTCCCCAGTTTCCAATGACCCTCCCCGTTGAGCCGGGGGCTTTCACATCAGACTTAAGAAACCGCCTGCGCGCGCTTTACGCCCAATAATTCCGGACAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAGGTACCGTCAAGGTACCGCCCTGTTCGAACGGTACTTGTTCTTCCCTAACAACAGAGTTTTACGATCCGAAAACCTTCATCACTCACGCGGCGTTGCTCCGTCAGACTTTCGTCCATTGCGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGCCGATCACCCTCTCAGGTCGGCTACGCATCGTCGCCTTGGTGAGCCGTTACCTCACCAACTAGCTAATGCGCCGCGGGTCCATCTGTAAGTGGTAGCTAAAAGCCACCTTTTATAATTGAACCATGCGGTTCAATCAAGCATCCGGTATTAGCCCCGTTTCCCGGAGTTATCCCAGTCTTACAGGCAGGTTACCCACGTGTTACTCACCCGTCCGCCGCTAACCTAAGGGAGCAAGCTCCCGTCGGTTCGCTCGACTTGCATGTATTAGGCACGCCGCCAGCGTTCGTCCTGAGCCTATACAAAAATCTATGGGTCCGATGTCGGATATTGGGCGTAAGCGCGCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGGAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAGAGGGTTTCCGCCCTTTAGTGCTGCAGCAAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTCTGACACCCCTAGAGATAGGGCTTCCCCTTCGGGGGCAGAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGCAGAACAAAGGGCAGCGAAGCCGCGAGGCTAAGCCAATCCCACAAATCTGCTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTGGAGCCAGCCGCCGAAGTGGGACAGATGATTGGGGTGAGCTAAAAGGAGTAGCCACAGCTAGTAAGGAGGTAGGCTTTCGCGCTCAGCGTCAGT TACAGACCAGAGAGTCGCCTTCGCCACTGGTGTTCCTCCACATCTCTACGCATTTCACCGCTACACGTGGAATTCCACTCTCCTCTTCTGCACTCAAGTTCCCCAGTTTCCAATGACCCTCCCCGTTGAGCCGGGGGCTTTCACATCAGACTTAAGAAACCGCCTGCGCGCGCTTTACGCCCAATAATTCCGGACAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAGGTACCGTCAAGGTACCGCCCTGTTCGAACGGTACTTGTTCTTCCCTAACAACAGAGTTTTACGATCCGAAAACCTTCATCACTCACGCGGCGTTGCTCCGTCAGACTTTCGTCCATTGCGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGCCGATCACCCTCTCAGGTCGGCTACGCATCGTCGCCTTGGTGAGCCGTTACCTCACCAACTAGCTAATGCGCCGCGGGTCCATCTGTAAGTGGTAGCTAAAAGCCACCTTTTATAATTGAACCATGCGGTTCAATCAAGCATCCGGTATTAGCCCCGTTTCCCGGAGTTATCCCAGTCTTACAGGCAGGTTACCCACGTGTTACTCACCCGTCCGCCGCTAACCTAAGGGAGCAAGCTCCCGTCGGTTCGCTCGACTTGCATGTATTAGGCACGCCGCCAGCGTTCGTCCTGAGCCTATACAAAAATCTAT
Bacillus Bacillus paralicheniformisparalicheniformis JJY12JJY12 -8 -8
GATGCGGATGTCGGATATTGGGCGTAAGCGCGCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGGAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAGAGGGTTTCCGCCCTTTAGTGCTGCAGCAAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTCTGACACCCCTAGAGATAGGGCTTCCCCTTCGGGGGCAGAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGCAGAACAAAGGGCAGCGAAGCCGCGAGGCTAAGCCAATCCCACAAATCTGCTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTGGAGCCAGCCGCCGAAGGTGGACAGATGATTGGGGTGAAGTCGTACGAAAAGCACACACAAAAGGATAAGTCTTTCGCGCCTCAGCGTCAGTTACAGACCAGAGAGTCGCCTTCGCCACTGGTGTTCCTCCACATCTCTACGCATTTCACCGCTACACGTGGAATTCCACTCTCCTCTTCTGCACTCAAGTTCCCCAGTTTCCAATGACCCTCCCCGGTTGAGCCGGGGGCTTTCACATCAGACTTAAGAAACCGCCTGCGCGCGCTTTACGCCCAATAATTCCGGACAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAGGTACCGTCAAGGCACCGCCCTGTTCGAACGGTACTTGTTCTTCCCTAACAACAGAGTTTTACGATCCGAAAACCTTCATCACTCACGCGGCGTTGCTCCGTCAGACTTTCGTCCATTGCGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGCCGATCACCCTCTCAGGTCGGCTACGCATCGTCGCCTTGGTGAGCCGTTACCTCACCAACTAGCTAATGCGCCGCGGGTCCATCTGTAAGTGGTAGCTAAAAGCCACCTTTTATAATTGAACCATGCGGTTCAATCAAGCATCCGGTATTAGCCCCGGTTTCCCGGAGTTATCCCAGTCTTACAGGCAGGTTACCCACGTGTTACTCACCCGTCCGCCGCTAACCTAAGGGAGCAAGCTCCCGTCGGTTCGCTCGACTTGCATGTATTAGGCACGCCGCCAGCGTTCGTCATGAGCCTGATCACCAACTCCCTGATGCGGATGTCGGATATTGGGCGTAAGCGCGCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGGAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAGAGGGTTTCCGCCCTTTAGTGCTGCAGCAAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTCTGACACCCCTAGAGATAGGGCTTCCCCTTCGGGGGCAGAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGCAGAACAAAGGGCAGCGAAGCCGCGAGGCTAAGCCAATCCCACAAATCTGCTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTGGAGCCAGCCGCCGAAGGTGGACAGATGATTGGGGTGAAGTCGTACGAAAAGCACACACAAAAGGATAAGTCTTTCGCGCCTCAGCGTCAGTTAC AGACCAGAGAGTCGCCTTCGCCACTGGTGTTCCTCCACATCTCTACGCATTTCACCGCTACACGTGGAATTCCACTCTCCTCTTCTGCACTCAAGTTCCCCAGTTTCCAATGACCCTCCCCGGTTGAGCCGGGGGCTTTCACATCAGACTTAAGAAACCGCCTGCGCGCGCTTTACGCCCAATAATTCCGGACAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAGGTACCGTCAAGGCACCGCCCTGTTCGAACGGTACTTGTTCTTCCCTAACAACAGAGTTTTACGATCCGAAAACCTTCATCACTCACGCGGCGTTGCTCCGTCAGACTTTCGTCCATTGCGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGCCGATCACCCTCTCAGGTCGGCTACGCATCGTCGCCTTGGTGAGCCGTTACCTCACCAACTAGCTAATGCGCCGCGGGTCCATCTGTAAGTGGTAGCTAAAAGCCACCTTTTATAATTGAACCATGCGGTTCAATCAAGCATCCGGTATTAGCCCCGGTTTCCCGGAGTTATCCCAGTCTTACAGGCAGGTTACCCACGTGTTACTCACCCGTCCGCCGCTAACCTAAGGGAGCAAGCTCCCGTCGGTTCGCTCGACTTGCATGTATTAGGCACGCCGCCAGCGTTCGTCATGAGCCTGATCACCAACTCCCT
Bacillus Bacillus sonorensissonorensis JJY13JJY13 -1 -One
GAAGTACGAGTCGGATATTGGGCGTAAGCGCGCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGGAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAGAGGGTTTCCGCCCTTTAGTGCTGCAGCAAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTCTGACACCCCTAGAGATAGGGCTTCCCCTTCGGGGGCAGAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGCAGAACAAAGGGCAGCGAAGCCGCGAGGCTAAGCCAATCCCACAAATCTGCTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTGGAGCCAGCCGCCGAAGTGGGACAGATGATTGGGGTGAGTCTCGGGGAGATGTCACACAGTCTAAAAGGATCAGCTTTCGCGCTCAGCGTCAGTTACAGACCAGAGAGTCGCCTTCGCCACTGGTGTTCCTCCACATCTCTACGCATTTCACCGCTACACGTGGAATTCCACTCTCCTCTTCTGCACTCAAGTTCCCCAGTTTCCAATGACCCTCCCCGGTTGAGCCGGGGGCTTTCACATCAGACTTAAGAAACCGCCTGCGCGCGCTTTACGCCCAATAATTCCGGACAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAGGTACCGTCAAGGCACCGCCCTGTTCGAACGGTACTTGTTCTTCCCTAACAACAGAGTTTTACGATCCGAAAACCTTCATCACTCACGCGGCGTTGCTCCGTCAGACTTTCGTCCATTGCGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGCCGATCACCCTCTCAGGTCGGCTACGCATCGTCGCCTTGGTGAGCCGTTACCTCACCAACTAGCTAATGCGCCGCGGGTCCATCTGTAAGTGGTAGCTAAAAGCCACCTTTTATAATTGAACCATGCGGTTCAATCAAGCATCCGGTATTAGCCCCGGTTTCCCGGAGTTATCCCAGTCTTACAGGCAGGTTACCCACGTGTTACTCACCCGTCCGCCGCTAACCTAAGGGAGCAAGCTCCCGTCGGTTCGCTCGACTTGCATGTATTAGGCACGCCGCCAGCGTTCGTCATGAGCCTAATCCAAAATCTCTAGAAGTACGAGTCGGATATTGGGCGTAAGCGCGCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGGAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAGAGGGTTTCCGCCCTTTAGTGCTGCAGCAAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTCTGACACCCCTAGAGATAGGGCTTCCCCTTCGGGGGCAGAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGCAGAACAAAGGGCAGCGAAGCCGCGAGGCTAAGCCAATCCCACAAATCTGCTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTGGAGCCAGCCGCCGAAGTGGGACAGATGATTGGGGTGAGTCTCGGGGAGATGTCACACAGTCTAAAAGGATCAGCTTTCGCGCTCAGCGTCAGT TACAGACCAGAGAGTCGCCTTCGCCACTGGTGTTCCTCCACATCTCTACGCATTTCACCGCTACACGTGGAATTCCACTCTCCTCTTCTGCACTCAAGTTCCCCAGTTTCCAATGACCCTCCCCGGTTGAGCCGGGGGCTTTCACATCAGACTTAAGAAACCGCCTGCGCGCGCTTTACGCCCAATAATTCCGGACAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAGGTACCGTCAAGGCACCGCCCTGTTCGAACGGTACTTGTTCTTCCCTAACAACAGAGTTTTACGATCCGAAAACCTTCATCACTCACGCGGCGTTGCTCCGTCAGACTTTCGTCCATTGCGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGCCGATCACCCTCTCAGGTCGGCTACGCATCGTCGCCTTGGTGAGCCGTTACCTCACCAACTAGCTAATGCGCCGCGGGTCCATCTGTAAGTGGTAGCTAAAAGCCACCTTTTATAATTGAACCATGCGGTTCAATCAAGCATCCGGTATTAGCCCCGGTTTCCCGGAGTTATCCCAGTCTTACAGGCAGGTTACCCACGTGTTACTCACCCGTCCGCCGCTAACCTAAGGGAGCAAGCTCCCGTCGGTTCGCTCGACTTGCATGTATTAGGCACGCCGCCAGCGTTCGTCATGAGCCTAATCCAAAATCTCTA
Bacillus Bacillus sonorensissonorensis JJY13JJY13 -3 -3
GACGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAGCGAACCGACGGGAGCTTGCTCCCTTAGGTTAGCGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCTGTAAGACTGGGATAACTCCGGGAACCGGGGCTAATACCGGATGCTTGATTGAACCGCATGGTTCAATTATAAAAGGTGGCTTTTAGCTACCACTTACAGATGGACCCGCGGCGCATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTTTCGGATCGTAAAACTCTGTTGTTAGGGAAGAACAAGTACCGTTCGAACAGGGCGGTACCTTGACGGTACCTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGGAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAGAGGGTTTCCGCCCTTTAGTGCTGCAGCAAACGCATTAAACTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTCTGACACCCCTAGAGATAGGGCTTCCCCTTCGGGGGCAGAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGCAGAACAAAGGGCAGCGAAGCCGCGAGGCTAAGCCAATCCCACAAATCTGCTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTGGAGCCAGCCGCCGAAAGTGGGACAGATGATTGGGGTGAAGACGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAGCGAACCGACGGGAGCTTGCTCCCTTAGGTTAGCGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCTGTAAGACTGGGATAACTCCGGGAACCGGGGCTAATACCGGATGCTTGATTGAACCGCATGGTTCAATTATAAAAGGTGGCTTTTAGCTACCACTTACAGATGGACCCGCGGCGCATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTTTCGGATCGTAAAACTCTGTTGTTAGGGAAGAACAAGTACCGTTCGAACAGGGCGGTACCTTGACGGTACCTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGGAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAGAGGGTTTCCGCCCTTTAGTGCTGCAGCAAACGCATTAAACTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTCTGACACCCCTAGAGATAGGG CTTCCCCTTCGGGGGCAGAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGCAGAACAAAGGGCAGCGAAGCCGCGAGGCTAAGCCAATCCCACAAATCTGCTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTGGAGCCAGCCGCCGAAAGTGGGACAGATGATTGGGGTGAA
Bacillus Bacillus sonorensissonorensis JJY13JJY13 -8 -8
GACGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAGCGAACCGACGGGAGCTTGCTCCCTTAGGTTAGCGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCTGTAAGACTGGGATAACTCCGGGAAACCGGGGCTAATACCGGATGCTTGATTGAACCGCATGGTTCAATTATAAAAGGTGGCTTTTAGCTACCACTTACAGATGGACCCGCGGCGCATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTTTCGGATCGTAAAACTCTGTTGTTAGGGAAGAACAAGTACCGTTCGAACAGGGCGGTACCTTGACGGTACCTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGGAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAGAGGGTTTCCGCCCTTTAGTGCTGCAGCAAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTCTGACACCCCTAGAGATAGGGCTTCCCCTTCGGGGGCAGAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGCAGAACAAAGGGCAGCGAAGCCGCGAGGCTAAGCCAATCCCACAAATCTGCTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTGGAGCCAGCCGCCGAAAGTGGGACAGATGATTGGGGTGGACGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAGCGAACCGACGGGAGCTTGCTCCCTTAGGTTAGCGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCTGTAAGACTGGGATAACTCCGGGAAACCGGGGCTAATACCGGATGCTTGATTGAACCGCATGGTTCAATTATAAAAGGTGGCTTTTAGCTACCACTTACAGATGGACCCGCGGCGCATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTTTCGGATCGTAAAACTCTGTTGTTAGGGAAGAACAAGTACCGTTCGAACAGGGCGGTACCTTGACGGTACCTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGGAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAGAGGGTTTCCGCCCTTTAGTGCTGCAGCAAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTCTGACACCCCTAGAGATA GGGCTTCCCCTTCGGGGGCAGAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGCAGAACAAAGGGCAGCGAAGCCGCGAGGCTAAGCCAATCCCACAAATCTGCTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTGGAGCCAGCCGCCGAAAGTGGGACAGATGATTGGGGTG
도 2는 본 발명에서 분리된 바실러스속 균주 5종의 당 발효능 평가 결과를 나타낸 도면으로, 균주 5종이 도면에 기재된 각 당류를 발효하여 해당 균주의 에너지원으로 이용할 수 있는 능력이 있는지 평가한 결과를 나타낸 것이다.FIG. 2 is a graph showing the results of evaluating the sugar-active activity of five strains of Bacillus sp. Isolated in the present invention. Five strains of the strains were fermented to show the ability to utilize the saccharides as an energy source .
도 2에 도시된 바와 같이, 균주 5종 각각이 많은 종류의 당에 대하여 반응하는 것을 알 수 있다. 즉, 바실러스 혼합 조성물을 구성하는 바실러스 소노렌시스(Bacillus sonorensis) JJY12-3, 바실러스 파라리체니포르미스(Bacillus paralicheniformis) JJY12-8, 바실러스 소노렌시스(Bacillus sonorensis) JJY13-1, 바실러스 소노렌시스(Bacillus sonorensis) JJY13-3, 바실러스 소노렌시스(Bacillus sonorensis) JJY13-8의 당 발효능에 대한 효과가 있는 것을 알 수 있다. 특히 바실러스 소노렌시스 JJY12-3은 Rhamnosus(15)와 Salicin(26) 발효능이 약한 반면 Potassium gluconate(47) 발효능이 높은 것을 확인할 수 있으며, 바실러스 소노렌시스 JJY13-1은 48시간 발효에서의 D-Xylose(6)과 Mteil-alphaD-manopyranoside(20), D-Rafinose(35)의 발효능이 높은 것을 알 수 있다. 또한 바실러스 소노렌시스 JJY13-1은 48시간 발효에서의 D-Xylose(6) 발효능은 높지만 Mteil-alphaD-manopyranoside(20)에 대한 발효능은 나타나지 않으며, 24시간 반응에서의 D-Lactose(29)와 D-Rafinose(35)의 발효능 또한 낮은 것을 알 수 있으며, 마지막으로 바실러스 소노렌시스 JJY13-8은 D-Xylose(6)와 D-Rafinose(35)의 발효능이 약하고 Mteil-alphaD-manopyranoside(20)의 발효능이 없는 것을 알 수 있다. 이와 같이, 16S 염기서열 동정에 의하여 같은 종(바실러스 소노렌시스)으로 분류된 균주들에 대하여 서로 다른 당 발효능이 존재함을 확인함으로써, 본 발명의 바실러스 5종이 서로 다른 균주임을 확인할 수 있다.As shown in Fig. 2, it can be seen that each of the five strains reacts with many kinds of sugars. That is, the Bacillus sonorensis JJY12-3, Bacillus paralicheniformis JJY12-8, Bacillus sonorensis JJY13-1, Bacillus sonorensis, ( Bacillus sonorensis ) JJY13-3, and Bacillus sonorensis JJY13-8. In particular, Bacillus thuringiensis JJY12-3 has a weak potency against Rhamnosus (15) and Salicin (26) while Potassium gluconate (47) has a high efficacy. D-Xylose (6), Mteil-alphaD-manopyranoside (20) and D-Rafinose (35) have high efficacy. In addition, Bacillus thuringiensis JJY13-1 showed high efficacy of D-Xylose (6) in fermentation for 48 hours, but no efficacy against Mteil-alphaD-manopyranoside (20) ) And D-Rafinose (35) were also found to be low. Finally, Bacillus thuringiensis JJY13-8 showed weak efficacy of D-Xylose (6) and D-Rafinose (35) and manopyranoside (20). Thus, by confirming the presence of different sugar-reducing activities on the strains classified as the same species (Bacillus thuringiensis) by the identification of the 16S nucleotide sequence, it can be confirmed that the five strains of Bacillus of the present invention are different strains.
도 3은 본 발명의 바실러스 혼합 조성물 섭취에 의한 체중 감소 효과를 나타낸 도면이고, 도 4는 본 발명의 바실러스 혼합 조성물 섭취에 의한 지방조직의 무게 감소 효과를 나타낸 도면이다.FIG. 3 is a graph showing the effect of the present invention on body weight reduction by ingesting a Bacillus mixed composition, and FIG. 4 is a graph showing an effect of weight reduction of adipose tissue by ingesting the Bacillus mixed composition of the present invention.
도 3과 도 4은 효창실험동물을 통해 구입한 6주령 C57BL/6 웅성 마우스를 2주간 예비 사육한 후, 무작위로 4그룹으로 나누어 실험한 것으로, 정상식이 그룹, 고지방식이 그룹, 고지방식이+VSL#3 그룹 및 고지방식이+바실러스 그룹으로 나누어 실험한 것이다(n=10). 여기서 n은 마우스의 개체수를 의미할 수 있다.FIGS. 3 and 4 show that 6-week-old C57BL / 6 male mice purchased from Hyochang experimental animals were preliminarily raised for 2 weeks and then randomly divided into 4 groups. + VSL # 3 group and highland method were divided into + Bacillus group (n = 10). Where n can refer to the number of mice.
여기서, 정상식이 그룹은 정상사료(2018S, Harlan Laboratories, Indianapolis, USA)를 섭취한 그룹으로 식이대조군을 의미하고, 고지방식이 그룹은 60%kcal 고지방식이(D12492, Research diets, New Brunswick, USA)를 섭취한 그룹으로 음성대조군(고지방식이 섭취에 의해 비만, 당뇨 및 지방간 유발)을 의미하며, 고지방식이+VSL#3 그룹은 고지방식이를 섭취하는 동시에 시판되는 복합 프로바이오틱스인 VSL#3 (5X108 CFU)를 섭취한 그룹으로 복합 프로바이오틱스 대조군을 의미하고, 고지방식이+바실러스 그룹은 고지방식이를 섭취하는 동시에 본 발명의 바실러스 혼합 조성물을 섭취한 그룹으로 바실러스 프로바이오틱스 실험군을 의미한다. 유의성 검증은 unpaired t test로 실시하였고, 각 유의성 수준은 "고지방식이" 그룹과 "고지방식이+바실러스" 그룹 간에는 *(p < 0.05), **(p < 0.01), ***(p < 0.001)로 표기하였으며, "정상식이" 그룹과 "고지방식이" 그룹 간에는 ###(p < 0.001)으로 표기하였다.The dietary control group was a 60% kcal diet (D12492, Research diets, New Brunswick, USA), and the control diet group was a group fed with normal diet (2018S, Harlan Laboratories, Indianapolis, USA) ), A negative control group (obesity, diabetes and fatty liver induced by ingestion by a high fat diet), and a high fat diet + VSL # 3 group consumes a high fat diet while simultaneously consuming a commercially available complex probiotic VSL # 3 (5 × 10 8 CFU). The term "high-fat diet + bacillus group" refers to a group of bacillus probiotics in which the high fat diet method is ingested and the bacillus mixed composition of the present invention is ingested. ( P <0.05), ** ( p <0.01), and *** ( p <0.01) between the high-fat diet group and the high fat diet + bacillus group <0.001), and between "normal diet" group and "high fat diet" group as ### ( p <0.001).
도 3과 도 4를 통해 알 수 있듯이, 본 발명의 바실러스 혼합 조성물을 섭취한 그룹(고지방식이+바실러스 그룹)이 바실러스 혼합 조성물의 섭취로 인하여 고지방식이 그룹과 고지방식이+VSL#3 그룹에 비해 체중 증가가 억제되어 체중 감소 효과를 갖는 것을 알 수 있으며, 특히 이러한 체중 감소 효과가 지방 조직의 무게 감소 효과에 기인함을 알 수 있다.As can be seen from FIGS. 3 and 4, when the group (high-fat diet + bacillus group) ingested the bacillus mixed composition of the present invention was consumed in the high fat diet group and the high fat diet + VSL # 3 group And weight reduction effect was suppressed. In particular, it can be understood that the weight reduction effect is due to the weight reduction effect of the fat tissue.
도 5는 본 발명의 바실러스 혼합 조성물 섭취에 의한 내당능 증대 효과를 나타낸 도면이고, 도 6은 본 발명의 바실러스 혼합 조성물 섭취에 의한 혈중 인슐린 감소 효과를 나타낸 도면이다.FIG. 5 is a graph showing an effect of increasing the glucose tolerance by ingesting a Bacillus mixed composition of the present invention, and FIG. 6 is a graph showing an effect of reducing the blood insulin level by ingesting the Bacillus mixed composition of the present invention.
도 5는 내당능 검사를 위하여 14주동안 각 시료를 섭취한 마우스를 대상으로 16시간 금식을 시행한 후, 2g/kg 포도당을 복강주사하고 0, 15, 30, 60, 90, 120분 후에 혈당측정기(GlucoDr Auto AGM-4000, Allmedicus, Anyang, Republic of Korea)를 사용하여 혈당수치를 측정한 것이고, 도 6은 혈중 인슐린 농도는 각 실험군을 대상으로 4시간 금식을 시행한 후 채혈한 혈액을 30분간 응고시킨 후 2,000g에서 30분간 원심분리하여 혈청을 얻고, Ultra Sensitive Mouse Insulin ELISA kit(Morinaga Institute of biological science Inc., Yokohama, Japan)를 이용하여 측정한 것이다.FIG. 5 is a graph showing the results of fasting for 16 hours in mice in which each sample was taken for 14 weeks for the glucose tolerance test. After 2, 4, 6, 6, the blood insulin concentration was measured by fasting for 4 hours in each experimental group, and the blood collected was counted for 30 minutes in each test group (GlucoDr Auto AGM-4000, Allmedicus, Anyang, Republic of Korea) After coagulation, the cells were centrifuged at 2,000 g for 30 minutes to obtain serum, which was measured using an Ultra Sensitive Mouse Insulin ELISA kit (Morinaga Institute of Biological Science Inc., Yokohama, Japan).
여기서, 정상식이 그룹은 정상사료(2018S, Harlan Laboratories, Indianapolis, USA)를 섭취한 그룹으로 식이대조군을 의미하고, 고지방식이 그룹은 60%kcal 고지방식이(D12492, Research diets, New Brunswick, USA)를 섭취한 그룹으로 음성대조군(고지방식이 섭취에 의해 비만, 당뇨 및 지방간 유발)을 의미하며, 고지방식이+VSL#3 그룹은 고지방식이를 섭취하는 동시에 시판되는 복합 프로바이오틱스인 VSL#3 (5X108 CFU)를 섭취한 그룹으로 복합 프로바이오틱스 대조군을 의미하고, 고지방식이+바실러스 그룹은 고지방식이를 섭취하는 동시에 본 발명의 바실러스 혼합 조성물을 섭취한 그룹으로 바실러스 프로바이오틱스 실험군을 의미한다. 유의성 검증은 unpaired t test로 실시하였고, 각 유의성 수준은 "고지방식이" 그룹과 "고지방식이+바실러스" 그룹 간에는 *(p < 0.05), **(p < 0.01)로 표기하였으며, "정상식이" 그룹과 "고지방식이" 그룹 간에는 #(p < 0.05), ###(p < 0.001)으로 표기하였다.The dietary control group was a 60% kcal diet (D12492, Research diets, New Brunswick, USA), and the control diet group was a group fed with normal diet (2018S, Harlan Laboratories, Indianapolis, USA) ), A negative control group (obesity, diabetes and fatty liver induced by ingestion by a high fat diet), and a high fat diet + VSL # 3 group consumes a high fat diet while simultaneously consuming a commercially available complex probiotic VSL # 3 (5X108 CFU). The term "Bacillus probiotics" refers to a group of high-fat diet + bacillus groups that consume a high fat diet while consuming the bacillus mixed composition of the present invention. The significance test was performed with unpaired t test. The significance level was indicated as * ( p <0.05), ** ( p <0.01) between "highland" and "highland + bacillus" ( P <0.05) and ### ( p <0.001) between the "group" and "highland" groups.
도 5와 도 6을 통해 알 수 있듯이, 본 발명의 바실러스 혼합 조성물을 섭취한 그룹(고지방식이+바실러스 그룹)이 바실러스 혼합 조성물의 섭취로 인하여 고지방식이 그룹과 고지방식이+VSL#3 그룹에 비해 내당능이 증대되고 혈중 인슐린 수치를 감소시키는 효과를 나타냄을 확인하였으며, 이에 따라 인슐린 감수성 촉진 효과가 있는 것을 알 수 있다.As shown in FIG. 5 and FIG. 6, when the group (high-fat diet + bacillus group) ingested the bacillus mixed composition of the present invention consumed the bacillus mixed composition, And the insulin levels in the blood were decreased. As a result, it was found that the insulin sensitivity promoting effect was enhanced.
도 7은 본 발명의 바실러스 혼합 조성물 섭취에 의한 지방간 개선 효과를 나타낸 도면이다.FIG. 7 is a graph showing the effect of the present invention on the improvement of fatty liver by ingesting a Bacillus mixed composition. FIG.
도 7의 실험 방법은 14주동안 각 시료를 섭취한 대조군 및 실험군 마우스로부터 간조직을 적출하고, 변형 방지를 위하여 10% 포르말린 용액에 적?한 조직을 고정한 후 4시간동안 흐르는 물에 수세하며, 일반적인 조직의 탈수, 투명화 및 파라핀 침투과정을 자동 조직 처리장치(TP1020, Leica Biosystems, Wetzlar, Germany)기기를 이용하여 시행한다. 파라핀 블록의 제작과 냉각은 자동 포매장치(EG1160, Leica Biosystems)를 사용하고, 조직 슬라이드 제작을 위하여 회전식 미세박절기(RM2255, Leica Biosystems)를 사용하여 조직을 수직 방향으로 4μm 두께로 연속절편하여 슬라이드에 부착한다. 조직 슬라이드는 헤마톡실린으로 염색하고 1% 염산을 함유한 70% 에탄올에 침지하여 핵을 청화하고, 대조 염색을 위하여 에오진으로 염색한 다음 흐르는 물에 세척하고 탈수 및 투명화과정을 거친다.In the experimental method of FIG. 7, liver tissues were extracted from the control group and the experimental group mice for 14 weeks, and the tissues were fixed in a 10% formalin solution to prevent deformation, and then the tissues were washed with water for 4 hours, General tissue dehydration, transparency, and paraffin infiltration are performed using an automatic tissue processor (TP1020, Leica Biosystems, Wetzlar, Germany). The paraffin block was fabricated and cooled using an automatic embedding device (EG1160, Leica Biosystems), and the tissue was successively sectioned 4 μm thick vertically using a rotary micrometer (RM2255, Leica Biosystems) . The tissue slides are dyed with hematoxylin, immersed in 70% ethanol containing 1% hydrochloric acid to purify the nuclei, stained with eosin for contrast dyeing, washed in flowing water, and subjected to dehydration and clarification.
여기서, 정상식이 그룹은 정상사료(2018S, Harlan Laboratories, Indianapolis, USA)를 섭취한 그룹으로 식이대조군을 의미하고, 고지방식이 그룹은 60%kcal 고지방식이(D12492, Research diets, New Brunswick, USA)를 섭취한 그룹으로 음성대조군(고지방식이 섭취에 의해 비만, 당뇨 및 지방간 유발)을 의미하며, 고지방식이+VSL#3 그룹은 고지방식이를 섭취하는 동시에 시판되는 복합 프로바이오틱스인 VSL#3 (5X108 CFU)를 섭취한 그룹으로 복합 프로바이오틱스 대조군을 의미하고, 고지방식이+바실러스 그룹은 고지방식이를 섭취하는 동시에 본 발명의 바실러스 혼합 조성물을 섭취한 그룹으로 바실러스 프로바이오틱스 실험군을 의미한다. 유의성 검증은 unpaired t test로 실시하였고, 각 유의성 수준은 "고지방식이" 그룹과 "고지방식이+바실러스" 그룹 간에는 *(p < 0.05)로 표기하였다.The dietary control group was a 60% kcal diet (D12492, Research diets, New Brunswick, USA), and the control diet group was a group fed with normal diet (2018S, Harlan Laboratories, Indianapolis, USA) ), A negative control group (obesity, diabetes and fatty liver induced by ingestion by a high fat diet), and a high fat diet + VSL # 3 group consumes a high fat diet while simultaneously consuming a commercially available complex probiotic VSL # 3 (5X108 CFU). The term "Bacillus probiotics" refers to a group of high-fat diet + bacillus groups that consume a high fat diet while consuming the bacillus mixed composition of the present invention. Significance tests were performed with the unpaired t test, and the significance levels were marked as * ( p <0.05) between the "Highland" and "Highland + Bacillus" groups.
도 7을 통해 알 수 있듯이, 본 발명의 바실러스 혼합 조성물을 섭취한 그룹(고지방식이+바실러스 그룹)이 바실러스 혼합 조성물의 섭취로 인하여 고지방식이 그룹과 고지방식이+VSL#3 그룹에 비해 간지방 축적이 감소하여 지방간 개선 효과가 있는 것을 알 수 있으며, 이에 의해 간조직의 무게가 감소하는 것을 알 수 있다.As can be seen from FIG. 7, when the group (high-fat diet + bacillus group) consumed the bacillus mixed composition of the present invention, the high fat diet group and the high fat diet group were significantly It can be seen that the fat accumulation is reduced and the fatty liver improving effect is exhibited, thereby decreasing the weight of the liver tissue.
또한, 본 발명은 상기 바실러스 혼합 조성물, 상기 바실러스 혼합 조성물의 배양물, 상기 배양물의 농축액 및 건조물로 이루어지는 군에서 선택된 1종 이상을 유효성분으로 포함하는 프로바이오틱스 제제를 제공할 수도 있다.In addition, the present invention may provide a probiotic preparation comprising at least one selected from the group consisting of the Bacillus mixed composition, the culture of the Bacillus mixed composition, the concentrate of the culture, and the dried product as an active ingredient.
상기 프로바이오틱스 제제는 당업계에 공지된 방법에 따라 다양한 제형과 방법으로 제조 및 투여될 수 있다. 예를 들어, 본 발명의 바실러스 혼합 조성물, 상기 바실러스 혼합 조성물, 상기 바실러스 혼합 조성물의 배양물, 상기 배양물의 농축액 및 건조물은 약제학적 분야에서 통상적으로 사용되는 담체와 혼합하여 산제(powder), 액제(liquids and solutions), 정제(tablet), 캡슐(capsule), 시럽(syrup), 현탁제(suspension) 또는 과립제(granule) 등의 형태로 제조되어 투여될 수 있다. 상기 담체로는 예를 들어, 결합제, 활탁제, 붕해제, 부형제, 가용화제, 분산제, 안정화제, 현탁화제, 색소 및 향료 등일 수 있으나, 이에 제한되지 않는다. 또한, 투여 용량은 체내에서의 활성성분의 흡수도, 불활성률, 배설속도, 피투여자의 연령, 성별, 축종, 상태 및 질병의 중증 정도 등에 따라 적절히 선택할 수 있다.The probiotic agent can be prepared and administered in various formulations and methods according to methods known in the art. For example, the Bacillus mixed composition, the Bacillus mixed composition, the Bacillus mixed composition culture, the concentrated solution and the dried product of the present invention may be mixed with a carrier commonly used in the pharmaceutical field to prepare a powder, for example, in the form of liquids and solutions, tablets, capsules, syrups, suspensions or granules. Examples of the carrier include, but are not limited to, binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, coloring matters and fragrances. The administration dose can be appropriately selected depending on the degree of absorption of the active ingredient in the body, the inactivation rate, the excretion rate, the age, sex, strain, condition and severity of the disease.
또한, 본 발명은 상기 바실러스 혼합 조성물, 상기 바실러스 혼합 조성물의 배양물, 상기 배양물의 농축액 및 건조물로 이루어지는 군에서 선택된 1종 이상을 유효성분으로 함유하는 비만 예방 또는 개선용 건강기능식품 조성물을 제공한다.The present invention also provides a health functional food composition for preventing or ameliorating obesity comprising, as an active ingredient, at least one member selected from the group consisting of the Bacillus mixed composition, the culture of the Bacillus mixed composition, the concentrate of the culture and the dried product .
본 발명의 건강기능식품 조성물은 정제, 과립, 분말, 캅셀, 액상의 용액 및 환으로 이루어진 군으로부터 선택된 어느 하나의 제형으로 제조된 것일 수 있다. 본 발명에 따른 건강기능식품 조성물은 상기 바실러스 혼합 조성물, 상기 바실러스 혼합 조성물의 배양물, 상기 배양물의 농축액 및 건조물 등을 유효성분으로 포함시켜 분말제, 액제, 정제, 연질캅셀제, 과립제, 티백차, 인스턴트 차 또는 드링크제 등의 형태로 제형화될 수 있다. 유효 성분으로서의 바실러스 혼합 조성물의 함량은 그 사용 목적(예방 또는 개선용)에 따라 적합하게 결정될 수 있다. 또한, 본 발명에 따른 식품 조성물은 바실러스 혼합 조성물 이외에 본 발명이 목적으로 하는 주효과를 손상시키지 않는 범위 내에서 바람직하게는 주효과에 상승효과를 줄 수 있는 다른 성분을 함유하는 것도 무방하다. The health functional food composition of the present invention may be prepared in any one form selected from the group consisting of tablets, granules, powders, capsules, liquid solutions and rings. The health functional food composition according to the present invention can be used as an active ingredient in the form of powders, liquids, tablets, soft capsules, granules, tea bags, Instant tea or a drink or the like. The content of the Bacillus mixed composition as an active ingredient can be appropriately determined depending on the intended use (for prevention or improvement). In addition, the food composition according to the present invention may contain, in addition to the Bacillus mixed composition, other ingredients which can give synergistic effects to the main effect, within the range not impairing the intended main effect of the present invention.
상기와 같은 형태로 제형화된 식품 조성물은 식품에 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 식품의 예로는 드링크제, 육류, 소시지, 빵, 비스킷, 떡, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 알코올 음료 및 비타민 복합제, 유제품 및 유가공 제품 등이 있으며, 통상적인 의미에서의 건강기능 식품은 모두 포함한다. The food composition formulated in this form can be added directly to the food or used together with other food or food ingredients, and can be suitably used according to conventional methods. Examples of food products include dairy products including drinks, meat, sausage, bread, biscuits, rice cakes, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, soups, , Dairy products and dairy products, and all health functional foods in the ordinary sense are included.
본 발명의 식품 조성물이 음료인 경우는 지시된 비율로 필수성분으로서 바실러스 혼합 조성물, 상기 바실러스 혼합 조성물의 배양물, 상기 배양물의 농축액 및 건조물을 함유하며, 그 밖의 음료 제조를 목적으로 사용되는 다른 성분에는 특별한 제한이 없으며 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상기한 천연 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스 등; 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상술한 것 이외의 향미제로서 천연 향미제 및 합성 향미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 ㎖당 일반적으로 약 1 내지 20 g, 바람직하게는 약 5 내지 12 g이다. When the food composition of the present invention is a beverage, it contains the Bacillus mixed composition, the culture of the Bacillus mixed composition, the concentrate and the dried product of the culture as essential ingredients at the indicated ratios, and the other ingredients And there may be added various flavoring agents or natural carbohydrates as an additional ingredient such as ordinary beverages. Examples of such natural carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors and synthetic flavors and the like can be used as the flavors other than those described above. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.
또한 본 발명의 식품 조성물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍 미제, 착색제 및 증진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에도 본 발명의 본 발명의 식품 조성물은 천연 과일 쥬스 및 과일 쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 그렇게 중요하진 않지만 본 발명의 바실러스 혼합 조성물 100 중량부 당 0.1 내지 약 20 중량부의 범위에서 선택되는 것이 일반적이다.In addition, the food composition of the present invention can be used as a food composition containing various nutrients, vitamins, minerals (electrolytes), synthetic flavors and flavors such as natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and its salts, , Organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the inventive food composition of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The ratio of such additives is not so important, but is generally selected in the range of 0.1 to about 20 parts by weight per 100 parts by weight of the Bacillus mixed composition of the present invention.
또한, 본 상기 바실러스 혼합 조성물, 상기 바실러스 혼합 조성물의 배양물, 상기 배양물의 농축액 및 건조물로 이루어지는 군에서 선택된 1종 이상을 유효성분으로 함유하는 비만 예방 또는 치료용 약학적 조성물을 제공한다.The present invention also provides a pharmaceutical composition for preventing or treating obesity, which comprises, as an active ingredient, at least one selected from the group consisting of the Bacillus mixed composition, the culture of the Bacillus mixed composition, the concentrate of the culture, and the dried product.
본 발명에서 사용되는 비만 예방 또는 치료용 약학적 조성물의 처리량은 약학적으로 유효한 양이어야 한다. 본 발명에서 사용되는 용어, "약학적으로 유효한 양"은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효 용량 수준은 개체 종류 및 중증도, 연령, 성별, 감염된 바이러스 종류, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출 비율, 치료 기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 유효량은 당업자에게 인식되어 있듯이 처리의 경로, 부형제의 사용 및 다른 약제와 함께 사용할 수 있는 가능성에 따라 변할 수 있다.The throughput of the pharmaceutical composition for preventing or treating obesity used in the present invention should be a pharmaceutically effective amount. As used herein, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and the effective dose level will vary depending on the species and severity, The type of virus being infected, the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and rate of release, the duration of the treatment, factors including co-administered drugs and other well known factors in the medical field. Effective amounts may vary depending on the route of treatment, the use of excipients, and the likelihood of use with other agents, as will be appreciated by those skilled in the art.
본 발명의 비만 예방 또는 치료용 약학적 조성물은 포유동물에 투여된 후 활성 성분의 신속, 지속 또는 지연된 방출을 제공할 수 있도록 당업계에 잘 알려진 방법을 사용하여 약학적 제형으로 제조될 수 있다. 제형의 제조에 있어서, 활성 성분을 담체와 함께 혼합 또는 희석하거나, 용기 형태의 담체 내에 봉입시키는 것이 바람직하다. The pharmaceutical composition for the prevention or treatment of obesity of the present invention may be manufactured into pharmaceutical formulations using methods well known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal. In the preparation of the formulations, it is preferred that the active ingredient is mixed with or diluted with the carrier, or enclosed in a carrier in the form of a container.
따라서, 본 발명의 비만 예방 또는 치료용 약학적 조성물은 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제 및 패치의 형태로 제형화하여 사용될 수 있고, 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 또는 희석제를 추가로 포함할 수 있다. Therefore, the pharmaceutical composition for preventing or treating obesity of the present invention can be formulated into oral formulations, external preparations and patches such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols according to a conventional method And may further comprise suitable carriers, excipients or diluents conventionally used in the manufacture of the compositions.
예를 들어, 본 발명의 약학 조성물에 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 광물유를 들 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제될 수 있다.Examples of carriers, excipients and diluents that can be included in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, it can be prepared using diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, surfactants and the like which are usually used.
또한, 본 명세서에 기재된 바실러스 혼합 조성물은 건강보조식품, 음료, 발효된 유제품을 포함하는 발효 식품 등과 같이 다양한 형태의 제품들에 함유될 수도 있다.In addition, the Bacillus mixed compositions described herein may be contained in various forms of products such as health supplements, beverages, fermented foods including fermented dairy products, and the like.
Claims (7)
Having obesity inhibitory effects, Bacillus Sono alkylene sheath (Bacillus sonorensis) JJY12-3 strain (Accession No: KCTC13405BP), Bacillus para-object miss formate (Bacillus paralicheniformis) JJY12-8 strain (Accession No: KCTC13406BP), Bacillus Sono alkylene cis (Bacillus sonorensis) JJY13-1 strain (Accession No: KCTC13407BP), Bacillus Sono alkylene sheath (Bacillus sonorensis) JJY13-3 strain (Accession No: KCTC13408BP), Bacillus Sono alkylene sheath (Bacillus sonorensis) JJY13-8 strain (Accession number: KCTC13409BP).
상기 바실러스 혼합 조성물은
당뇨 예방 효능 및 지방간 축적 억제 효능을 갖는 것을 특징으로 하는 바실러스 혼합 조성물.
The method according to claim 1,
The Bacillus mixed composition
A diabetic prophylactic effect and an anti-fat deposition effect.
A probiotic preparation comprising at least one selected from the group consisting of the Bacillus mixed composition of any one of claims 1 to 2, the culture of the Bacillus mixed composition, the concentrate of the culture and the dried product as an active ingredient.
A health functional food composition comprising, as an active ingredient, at least one selected from the group consisting of the Bacillus mixed composition of any one of claims 1 to 2, the culture of the Bacillus mixed composition, the concentrate of the culture and the dried product.
A pharmaceutical composition comprising, as an active ingredient, at least one selected from the group consisting of the Bacillus mixed composition of any one of claims 1 to 2, the culture of the Bacillus mixed composition, the concentrate of the culture and the dried product.
A beverage comprising at least one of the Bacillus mixed composition of any one of claims 1 to 2, the culture of the Bacillus mixed composition, the concentrate of the culture and the dried product as an active ingredient.
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