KR102630612B1 - Food Composition for Preventing or Improving Oral Disease or Colorectal Disease, Comprising a new Enterococcus faecalis strain and its whey fermentation product - Google Patents
Food Composition for Preventing or Improving Oral Disease or Colorectal Disease, Comprising a new Enterococcus faecalis strain and its whey fermentation product Download PDFInfo
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- KR102630612B1 KR102630612B1 KR1020210019288A KR20210019288A KR102630612B1 KR 102630612 B1 KR102630612 B1 KR 102630612B1 KR 1020210019288 A KR1020210019288 A KR 1020210019288A KR 20210019288 A KR20210019288 A KR 20210019288A KR 102630612 B1 KR102630612 B1 KR 102630612B1
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- A—HUMAN NECESSITIES
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Abstract
본 발명은 유청 발효 시 구강질환 또는 대장질환 개선 효능을 나타내는 수탁번호 KACC 81148BP의 엔테로코커스 패칼리스 (Enterococcus faecalis) M157 균주, 상기 균주로 바이오컨버전(bioconversion)된 유청 발효물을 포함하는 구강질환 또는 대장질환의 예방 또는 개선용 식품 조성물, 건강기능식품, 식품첨가제 조성물, 약학적 조성물, 의약외품 조성물 및 사료 조성물에 관한 것이다.
본 발명에서 엔테로코커스 패칼리스 M157 균주로 바이오컨버전된 유청 발효물은 치주조직의 총 부피 당 골 부피 증가, 골 부피 당 표면적 감소, 골 소실 수피 감소 효과를 나타내며 치주 조직 및 대장 조직에서 전염증 지표를 감소시키고 항염증 지표를 증가시키는 바, 구강 질환 및 대장 질환의 예방, 개선 또는 치료용으로 유용하게 사용될 수 있어 산업적 활용 가치가 높다.The present invention relates to the Enterococcus faecalis M157 strain with accession number KACC 81148BP, which shows efficacy in improving oral or colonic diseases during whey fermentation, and oral diseases or colonic diseases including whey fermentation product bioconverted with the strain. It relates to food compositions for preventing or improving diseases, health functional foods, food additive compositions, pharmaceutical compositions, quasi-drug compositions, and feed compositions.
In the present invention, the whey fermentation product bioconverted to Enterococcus faecalis M157 strain increases bone volume per total volume of periodontal tissue, decreases surface area per bone volume, reduces bone loss and bark, and exhibits pro-inflammatory indicators in periodontal tissue and colon tissue. As it reduces and increases anti-inflammatory indicators, it can be useful for preventing, improving or treating oral diseases and colon diseases, so it has high industrial value.
Description
본 발명은 유청 발효 시 구강질환 또는 대장질환 개선 효능을 나타내는 수탁번호 KACC 81148BP의 엔테로코커스 패칼리스 (Enterococcus faecalis) M157 균주, 상기 균주로 바이오컨버전(bioconversion)된 유청 발효물을 포함하는 구강질환 또는 대장질환의 예방 또는 개선용 식품 조성물, 식품 첨가제 조성물, 건강기능식품, 약학적 조성물, 의약외품 조성물 및 사료 조성물에 관한 것이다.The present invention relates to the Enterococcus faecalis M157 strain with accession number KACC 81148BP, which shows efficacy in improving oral or colonic diseases during whey fermentation, and oral diseases or colonic diseases including whey fermentation product bioconverted with the strain. It relates to food compositions for preventing or improving diseases, food additive compositions, health functional foods, pharmaceutical compositions, quasi-drug compositions, and feed compositions.
"유청(whey)"은 우유를 치즈로 가공할 때 형성되는 부산물로, 탈지유에 산 또는 응유효소를 첨가하여 생성되는 응고물, 즉 응유(curd)를 제외한 수용액을 의미한다. 종래에는 유청이 치즈를 생산함에 있어 단지 부산물로 여겨져 매년 다량 폐기되곤 하였으나, 최근에는 그 영양학적 가치를 인정받아 이의 활용도를 높이고 고부가 가치화를 이루고자 하는 노력이 점차 증가하고 있는 추세이다.“Whey” is a by-product formed when milk is processed into cheese, and refers to an aqueous solution excluding curds, which is a curd produced by adding acid or milk-curd enzymes to skim milk. Previously, whey was considered only a by-product of cheese production and was discarded in large quantities every year, but recently, its nutritional value has been recognized, and efforts to increase its utilization and achieve high added value are gradually increasing.
유청 단백질의 시장 규모는 2018년 기준, 약 74억 불(한화 약 9조 원)에 달하며, 2025년까지 8.1%의 연평균 성장률을 보일 것으로 예상된다. 유청 단백질은 생리 활성 펩타이드 생산 물질로 주목을 받고 있으며 다양한 고부가가치 산업 분야에서 사용될 가능성이 있어 이에 관한 연구가 활발히 이루어지고 있다.As of 2018, the whey protein market size is approximately $7.4 billion (approximately KRW 9 trillion) and is expected to show an average annual growth rate of 8.1% until 2025. Whey protein is attracting attention as a material for producing bioactive peptides, and research on it is being actively conducted as it has the potential to be used in various high value-added industries.
"바이오컨버전(bioconversion)" 기술은 생물전환 기술이라고도 불리우며, 미생물 등이 가지고 있는 효소적 기능을 이용하여 전구물질로부터 보다 부가가치가 높은 유용물질을 생산하는 과정을 의미한다. 즉, 미생물 발효 또는 효소 처리 등의 생물학적 방법을 통해 생리활성 물질의 구조적 변화를 유도함으로써 특정 유효성분의 함량을 증가시키거나, 흡수율을 개선하거나, 새로운 유효성분의 생성을 유도하는 기술로서 최근 각광받고 있다. 그러나, 이러한 기술에 대한 연구에도 불구하고 실제 최종 제품의 생산으로 이어지는 것은 미미한 실정이었다.“Bioconversion” technology, also called bioconversion technology, refers to the process of producing useful substances with higher added value from precursor materials using the enzymatic functions of microorganisms, etc. In other words, it has recently been in the spotlight as a technology that increases the content of specific active ingredients, improves absorption rate, or induces the creation of new active ingredients by inducing structural changes in bioactive substances through biological methods such as microbial fermentation or enzyme treatment. there is. However, despite research into these technologies, little has been achieved in producing actual final products.
한편, 구강질환은 치아를 포함한 구강 내에서 발생할 수 있는 모든 질환을 일컫는 것으로, 흔히 발생하는 구강질환으로는 대표적으로 치아 우식증(dental caries), 즉 충치와 치주염(periodontitis), 치은염(gingivitis) 등의 치주질환이 있을 수 있다.Meanwhile, oral diseases refer to all diseases that can occur in the oral cavity, including teeth. Common oral diseases include dental caries, cavities, periodontitis, and gingivitis. You may have periodontal disease.
이러한 구강질환 또는 치주질환은 국내외를 불문하고 성인 인구에 대해 가장 흔하게 발생할 수 있는 질환에 해당할 뿐만 아니라, 개인 의료 및 사회적인 관리 체계 면에서도 상당한 비용이 요구되는, 공중 보건의 관심질환에 해당하며, 특히 치주 질환은 전신 만성 질병과 관련이 있어 대장 질환과의 상관성이 대두되며 구강-장내 균총에 관한 활발한 연구가 이루어지고 있다.These oral diseases or periodontal diseases are not only the most common diseases in the adult population, both at home and abroad, but they are also diseases of public health concern that require significant costs in terms of personal medical and social management systems. In particular, periodontal disease is related to systemic chronic diseases, so its correlation with colonic disease is emerging, and active research is being conducted on oral-intestinal flora.
국내에서 구강 건강을 위한 건강기능식품으로 제품에 활용된 프로바이오틱스에는 Lactobacillus reuteri, L. salivarius, L. plantarum, Weisella cibaria 등이 있으나, 국내에서 치주염 개선 혹은 구강 및 대장 건강을 위한 유청발효 산물 제품 개발은 부족한 실정이다. In Korea, probiotics used in products as health functional foods for oral health include Lactobacillus reuteri, L. salivarius, L. plantarum, and Weisella cibaria. However, in Korea, there has been no development of whey fermentation product products for improving periodontitis or oral and colon health. There is a shortage.
이에 본 발명의 발명자는 종래에 대량 폐기되는 유청의 활용 가능성을 재고함과 동시에, 유청으로부터 보다 부가가치가 높은 구강질환 또는 대장질환 개선 물질을 생산하고자 예의 노력한 결과, 유청 분말에 대해 원유로부터 분리된 엔테로코커스 패칼리스 M157 균주 (수탁번호 : KACC 81148BP)를 접종하고 배양시킨 유청 발효물이 바이오컨버전을 통해 구강질환 및 대장질환에 도움을 줄 수 있음을 확인하여 본 발명을 완성하였다. Accordingly, the inventor of the present invention reconsidered the possibility of utilizing whey, which was previously discarded in large quantities, and at the same time made diligent efforts to produce substances for improving oral or colonic diseases with higher added value from whey. The present invention was completed by confirming that the fermented whey product inoculated and cultured with the Coccus faecalis M157 strain (accession number: KACC 81148BP) can help oral and colonic diseases through bioconversion.
본 발명은 전술한 문제 및 이와 연관된 다른 문제를 해결하는 것을 목적으로 한다.The present invention aims to solve the above-described problems and other problems associated therewith.
본 발명의 일 예시적 목적은 엔테로코커스 패칼리스 (Enterococcus faecalis) 균주로 바이오컨버전 (bioconversion)된 유청 발효물을 유효성분으로 포함하는, 구강질환 또는 대장질환 예방 또는 개선용 식품 조성물을 제공하는 것이다.An exemplary object of the present invention is to provide a food composition for preventing or improving oral disease or colonic disease, which contains fermented whey bioconverted with Enterococcus faecalis strain as an active ingredient.
본 발명의 다른 예시적 목적은 엔테로코커스 패칼리스 (Enterococcus faecalis) 균주로 바이오컨버전 (bioconversion)된 유청 발효물을 유효성분으로 포함하는, 구강질환 또는 대장질환 예방 또는 개선용 건강기능식품을 제공하는 것이다.Another exemplary object of the present invention is to provide a health functional food for preventing or improving oral or colonic diseases, which contains fermented whey bioconverted with Enterococcus faecalis strain as an active ingredient. .
본 발명의 또 다른 예시적 목적은 엔테로코커스 패칼리스 (Enterococcus faecalis) 균주로 바이오컨버전 (bioconversion)된 유청 발효물을 유효성분으로 포함하는, 구강질환 또는 대장질환 예방 또는 개선용 식품 첨가제 조성물을 제공하는 것이다.Another exemplary object of the present invention is to provide a food additive composition for preventing or improving oral disease or colonic disease, comprising fermented whey bioconverted with Enterococcus faecalis strain as an active ingredient. will be.
본 발명의 또 다른 예시적 목적은 유청 발효 시 구강질환 또는 대장질환 개선 효능을 나타내는 수탁번호 KACC 81148BP의 엔테로코커스 패칼리스 M157 균주를 제공하는 것이다.Another exemplary object of the present invention is to provide an Enterococcus faecalis M157 strain with accession number KACC 81148BP that shows efficacy in improving oral or colonic diseases during whey fermentation.
본 발명의 또 다른 예시적 목적은 엔테로코커스 패칼리스 균주로 바이오컨버전된 유청 발효물을 유효성분으로 포함하는, 구강질환 또는 대장질환 예방 또는 치료용 약학적 조성물을 제공하는 것이다.Another exemplary object of the present invention is to provide a pharmaceutical composition for preventing or treating oral disease or colon disease, which contains fermented whey bioconverted with Enterococcus faecalis strain as an active ingredient.
본 발명의 또 다른 예시적 목적은 엔테로코커스 패칼리스 균주로 바이오컨버전된 유청 발효물을 유효성분으로 포함하는, 구강질환 또는 대장질환 예방 또는 개선용 의약외품 조성물을 제공하는 것이다.Another exemplary object of the present invention is to provide a quasi-drug composition for preventing or improving oral or colonic diseases, comprising fermented whey bioconverted to Enterococcus faecalis strains as an active ingredient.
본 발명의 또 다른 예시적 목적은 엔테로코커스 패칼리스 균주로 바이오컨버전된 유청 발효물을 유효성분으로 포함하는, 구강질환 또는 대장질환 예방 또는 개선용 사료 조성물을 제공하는 것이다.Another exemplary object of the present invention is to provide a feed composition for preventing or improving oral disease or colonic disease, comprising fermented whey bioconverted with Enterococcus faecalis strain as an active ingredient.
본 명세서에 개시된 발명의 기술적 사상에 따라 이루고자 하는 기술적 과제는 이상에서 언급한 문제점을 해결하기 위한 과제로 제한되지 않으며, 언급되지 않은 또 다른 과제는 아래의 기재로부터 통상의 기술자에게 명확하게 이해될 수 있을 것이다.The technical problem to be achieved according to the technical idea of the invention disclosed in this specification is not limited to the problem to solve the problems mentioned above, and other problems not mentioned can be clearly understood by those skilled in the art from the description below. There will be.
이를 구체적으로 설명하면 다음과 같다. 한편, 본 출원에서 개시된 각각의 설명 및 실시형태는 각각의 다른 설명 및 실시 형태에도 적용될 수 있다. 즉, 본 출원에서 개시된 다양한 요소들의 모든 조합이 본 출원의 범주에 속한다. 또한, 하기 기술된 구체적인 서술에 의하여 본 출원의 범주가 제한된다고 볼 수 없다.This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present application may also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in this application fall within the scope of this application. Additionally, the scope of the present application cannot be considered limited by the specific description described below.
상기 목적을 달성하기 위한 일 양태로서, 본 발명은 엔테로코커스 패칼리스 (Enterococcus faecalis, E. faecalis) 균주로 바이오컨버전 (bioconversion)된 유청 발효물을 유효성분으로 포함하는, 구강질환 또는 대장질환 예방 또는 개선용 식품 조성물을 제공한다.In one aspect for achieving the above object, the present invention is directed to preventing oral disease or colonic disease, comprising fermented whey bioconverted to Enterococcus faecalis (E. faecalis ) strain as an active ingredient, or A food composition for improvement is provided.
본 발명에 있어서, 상기 엔테로코커스 패칼리스 균주는 수탁번호 KACC 81148BP의 엔테로코커스 패칼리스(Enterococcus faecalis) M157 균주일 수 있으나, 이에 제한되지 않는다.In the present invention, the Enterococcus faecalis strain may be Enterococcus faecalis M157 strain with accession number KACC 81148BP, but is not limited thereto.
엔테로코커스 패칼리스(Enterococcus faecalis) M157 균주는 원유에서 분리되어 구강질환 및 대장질환 개선 효능을 나타내는 균주로, 2020년 11월 25일 자로 국립농업과학원 미생물은행(KACC)에 기탁되어 수탁번호 KACC 81148BP를 부여받았다.Enterococcus faecalis M157 strain is isolated from raw milk and shows efficacy in improving oral and colon diseases. It was deposited in the Microbial Bank of the National Academy of Agricultural Sciences (KACC) on November 25, 2020, and has accession number KACC 81148BP. It was granted.
본 발명에서 용어 "유청(whey)"은 우유를 치즈로 가공할 때 형성되는 부산물로, 탈지유에 산 또는 응유효소를 첨가하여 생성되는 응고물, 즉 응유(curd)를 제외한 수용액을 의미한다.In the present invention, the term "whey" is a by-product formed when milk is processed into cheese, and refers to an aqueous solution excluding curd, which is a coagulated product produced by adding acid or milk curd enzyme to skim milk.
본 발명에서 용어 "발효물"은 미생물을 이용한 효소적 또는 대사적 분해의 결과물을 의미하는 것으로, 구체적으로는 본 발명에서는 유청에 엔테로코커스 패칼리스 M157 균주를 접종한 후 배양한 산물을 의미할 수 있다. 또한 본 명세서 내에서 '발효물' 또는 '유청 발효물'은 '바이오컨버전 산물'과 혼용될 수 있다.In the present invention, the term "fermented product" refers to the result of enzymatic or metabolic degradation using microorganisms. Specifically, in the present invention, it may refer to a product obtained by inoculating whey with Enterococcus faecalis M157 strain and then culturing it. there is. Additionally, within this specification, ‘fermented product’ or ‘whey fermented product’ may be used interchangeably with ‘bioconversion product’.
본 발명에서 용어 "바이오컨버전(bioconversion)"은 생물전환이라고도 하며, 미생물이 가지고 있는 효소적 기능을 이용하여 전구물질로부터 보다 부가가치가 높은 유용물질을 생산하는 것을 의미한다. 즉, 미생물 발효 또는 효소 처리 등의 생물학적 방법을 통해 생리활성 물질의 구조적 변화를 유도하여 유효성분의 함량 증가, 흡수율 개선 및 새로운 유효성분의 생성을 유도할 수 있다. 본 발명에서 바이오컨버전 산물은 유청에 엔테로코커스 패칼리스 M157균주를 접종한 후 배양한 유청 발효물을 의미할 수 있으며, 상기 바이오컨버전 산물을 통해 구강질환 및 대장질환의 예방, 개선 또는 치료 효과를 달성할 수 있다. In the present invention, the term “bioconversion” is also referred to as bioconversion and refers to the production of useful substances with higher added value from precursors using the enzymatic functions of microorganisms. In other words, structural changes in bioactive substances can be induced through biological methods such as microbial fermentation or enzyme treatment, leading to increased content of active ingredients, improved absorption rate, and creation of new active ingredients. In the present invention, the bioconversion product may refer to a whey fermentation product cultured after inoculating whey with Enterococcus faecalis M157 strain, and the bioconversion product achieves the effect of preventing, improving, or treating oral diseases and colonic diseases. can do.
본 발명에 있어서, 유청 발효물은 엔테로코커스 패칼리스 균주로 바이오컨버전된 것일 수 있으며, 보다 구체적으로 수탁번호 KACC 81148BP의 엔테로코커스 패칼리스 M157 균주로 바이오컨버전된 것일 수 있다.In the present invention, the whey fermentation product may be bioconverted into an Enterococcus faecalis strain, and more specifically, may be bioconverted into an Enterococcus faecalis M157 strain with accession number KACC 81148BP.
본 발명에서 용어 "구강질환"은 치아를 포함하여 구강 내에서 발생할 수 있는 모든 질환을 의미하는 것으로, 구체적으로 구강 조직 내에서 파골세포 수 감소, 조골세포 수 증가, 해면골 소실 부피 감소, 치조골 부피 증가, 염증세포 수 감소 및 구강 유해세균에 대한 항균 효과로 인해 예방, 개선 및/또는 치료될 수 있는 모든 질환을 포함한다. 보다 구체적으로, 상기 구강질환은 치조골 파손(alveolar bone breakage), 치조골다공증(alveolar bone osteoporosis), 치조골연화증(alveolar bone osteomalacia), 치조골감소증(alveolar bone osteopenia), 치아 우식증(dental caries), 치은염(gingivitis) 및 치주염(periodontitis)으로 구성된 군으로부터 선택되는 하나 이상의 질환일 수 있으며, 이에 특별히 제한되는 것은 아니다.In the present invention, the term "oral disease" refers to all diseases that can occur in the oral cavity, including teeth. Specifically, in oral tissues, the number of osteoclasts decreases, the number of osteoblasts increases, the volume of cancellous bone is lost, and the alveolar bone volume increases. , includes all diseases that can be prevented, improved, and/or treated due to the reduction in the number of inflammatory cells and the antibacterial effect on harmful oral bacteria. More specifically, the oral diseases include alveolar bone breakage, alveolar bone osteoporosis, alveolar bone osteomalacia, alveolar bone osteopenia, dental caries, and gingivitis. ) and periodontitis, but is not particularly limited thereto.
본 발명에서 용어 "대장질환"은 대장에서 발생할 수 있는 모든 질환을 의미하는 것으로, 구체적으로 대장암, 대장용종, 과민성장증후군, 궤양성대장염 및 크론병, 결핵성장염, 감염성 설사 및 대장염, 혈관성장질환, 대장게실증으로 구성된 군으로부터 선택되는 하나 이상의 질환일 수 있으며, 이에 특별히 제한되는 것은 아니다.In the present invention, the term "colon disease" refers to all diseases that can occur in the colon, specifically colon cancer, colon polyps, irritable bowel syndrome, ulcerative colitis and Crohn's disease, tuberculous enteritis, infectious diarrhea and colitis, and vascular growth. It may be one or more diseases selected from the group consisting of disease and colonic diverticulosis, but is not particularly limited thereto.
본 발명에서 용어 "예방"은 본 발명 조성물의 투여로 구강질환 또는 대장질환과 관련된 증상을 억제 또는 지연시키는 모든 행위를 의미하며, 본 발명에서 용어 "개선"은 본 발명의 조성물을 적용하여 구강질환 또는 대장질환의 증상을 완화시키는 효과를 나타내는 것을 의미한다.In the present invention, the term "prevention" refers to all actions that suppress or delay symptoms related to oral disease or colon disease by administering the composition of the present invention, and the term "improvement" in the present invention refers to the treatment of oral disease by applying the composition of the present invention. Or, it means showing the effect of alleviating the symptoms of colon disease.
본 발명의 일 실시예에서는 치주염을 유발한 랫드에 대해 엔테로코커스 패칼리스 M157 균주에 의한 바이오컨버전 산물을 경구투여 및 치아도포한 결과, 바이오컨버전 산물을 투여한 군에서 치주 조직의 총 부피 당 골 부피 증가, 골 부피 당 표면적 감소, 골 소실 부피 감소로 인한 치주염 치료 효과가 현저히 우수함을 확인하였다. 또한 치주조직과 대장조직에서 전사체 및 단백질 발현량을 분석한 결과, 엔테로코커스 패칼리스 M157 균주에 의한 바이오컨버전 산물을 처리한 랫드에서 전염증 지표 발현량이 낮게 관찰되고, 항염증 및 항산화 지표 발현량이 높게 관찰되어, 치주 조직 및 대장 조직의 염증 개선 효과가 현저히 우수함을 확인하였다.In one embodiment of the present invention, as a result of oral administration and dental application of a bioconversion product by Enterococcus faecalis M157 strain to rats inducing periodontitis, the bone volume per total volume of periodontal tissue in the group administered the bioconversion product was It was confirmed that the treatment effect for periodontitis was significantly superior due to the increase in bone volume, decrease in surface area per bone volume, and decrease in bone loss volume. Additionally, as a result of analyzing transcript and protein expression levels in periodontal and colonic tissues, low expression levels of pro-inflammatory indicators were observed in rats treated with bioconversion products by Enterococcus faecalis M157 strain, and expression levels of anti-inflammatory and antioxidant indicators were low. It was observed to be high, confirming that the effect of improving inflammation in periodontal tissue and colon tissue was significantly excellent.
본 발명의 식품 조성물은 다양한 형태의 제형으로 제조될 수 있으며, 일반 약품과는 달리 식품을 원료로 하여 약품의 장기 복용 시 발생할 수 있는 부작용 등이 없는 장점이 있다. 본 발명의 식품 조성물은 어떠한 형태로도 제조될 수 있으며, 구체적으로는 정제, 캡슐, 환, 과립, 액상, 분말, 편상, 페이스트상, 시럽, 겔, 젤리, 바 등의 건강기능식품 제제류, 음료류, 껌류 및 캔디류로 구성된 군으로부터 선택되는 하나 이상의 제형일 수 있으나, 이에 특별히 제한되는 것은 아니다.The food composition of the present invention can be manufactured in various types of formulations, and unlike general drugs, it has the advantage of being made from food as a raw material and having no side effects that may occur when taking the drug for a long period of time. The food composition of the present invention can be manufactured in any form, and specifically, health functional food preparations such as tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, bars, etc. It may be one or more formulations selected from the group consisting of beverages, gums, and candies, but is not particularly limited thereto.
또한, 본 발명의 식품 조성물에는 그 유효성분 이외에 식품 첨가물이 포함될 수 있다. 식품 첨가물은 일반적으로 식품을 제조, 가공 또는 보존함에 있어 식품에 첨가되어 혼합되거나 침윤되는 물질로서 이해될 수 있으며, 식품과 함께 매일 그리고 장기간 섭취되므로 그 안전성이 보장되어야 한다. 상기 식품 첨가물은 기능 면에 있어서 감미제, 풍미제, 보존제, 유화제, 산미료, 점증제 등으로 구분되며, 본 발명의 식품 조성물이 달성하고자 하는 목적에 부합하는 한 특별히 제한되지 않는다. 또한, 본 발명의 식품 조성물은 상기 식품 첨가물 이외에 기능성과 영양 보충의 목적으로 당업계에 공지되고 식품 첨가물로서 안정성이 보장된 생리활성 물질 또는 미네랄류를 포함할 수 있다. 상기 생리활성 물질 또는 미네랄류는 본 발명의 식품 조성물이 달성하고자 하는 목적에 부합하는 한 특별히 제한되지 않는다.Additionally, the food composition of the present invention may contain food additives in addition to the active ingredients. Food additives can generally be understood as substances that are added to, mixed with, or infiltrated into food when manufacturing, processing, or preserving food. Since they are consumed daily and for a long period of time with food, their safety must be guaranteed. The food additives are classified into sweeteners, flavorants, preservatives, emulsifiers, acidulants, thickeners, etc. in terms of function, and are not particularly limited as long as they meet the purpose to be achieved by the food composition of the present invention. In addition, the food composition of the present invention may contain, in addition to the food additives, bioactive substances or minerals known in the art for the purpose of functionality and nutritional supplementation and whose safety is guaranteed as food additives. The physiologically active substances or minerals are not particularly limited as long as they meet the purpose to be achieved by the food composition of the present invention.
본 발명의 식품 조성물에는 전술한 바의 식품 첨가물이 제품 유형에 따라 그 첨가 목적을 달성할 수 있는 유효량으로 포함될 수 있으며, 본 발명의 식품 조성물에 포함될 수 있는 기타의 식품 첨가물과 관련하여서는 각국 식품공전이나 식품 첨가물 공전을 참조할 수 있다.The food composition of the present invention may contain the above-mentioned food additives in an effective amount to achieve the purpose of addition depending on the product type, and with respect to other food additives that may be included in the food composition of the present invention, each country's food code You can also refer to the Food Additives Code.
본 발명의 다른 양태로서, 본 발명은 엔테로코커스 패칼리스 (Enterococcus faecalis) 균주로 바이오컨버전 (bioconversion)된 유청 발효물을 유효성분으로 포함하는, 구강질환 또는 대장질환 예방 또는 개선용 건강기능식품을 제공한다.In another aspect of the present invention, the present invention provides a health functional food for preventing or improving oral disease or colon disease, comprising fermented whey bioconverted with Enterococcus faecalis strain as an active ingredient. do.
상기 용어 엔테로코커스 패칼리스 균주, 바이오컨버전, 유청, 발효물, 구강질환, 대장질환 및 예방은 전술한 바와 같다.The terms Enterococcus faecalis strain, bioconversion, whey, fermented product, oral disease, colon disease, and prevention are the same as described above.
본 발명에서 용어 "건강기능식품"이란, 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 말한다. 상기 "기능성" 은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건용도에 유용한 효과를 얻는 것을 의미한다. 본 발명의 건강기능식품은 당업계에서 통상적으로 사용되는 방법에 의하여 제조 가능하며, 제조시 당업계에서 통상적으로 첨가하는 원료 및 성분을 첨가하여 제조할 수 있다. 또한 상기 건강기능식품의 제형 또한 건강기능식품으로 인정되는 제형이면 제한없이 제조될 수 있다. In the present invention, the term “health functional food” refers to food manufactured and processed using raw materials or ingredients with functional properties useful to the human body. The term “functionality” refers to obtaining effects useful for health purposes, such as controlling nutrients or physiological effects on the structure and function of the human body. The health functional food of the present invention can be manufactured by a method commonly used in the art, and can be manufactured by adding raw materials and components commonly added in the art during production. Additionally, the formulation of the health functional food can also be manufactured without limitation as long as it is a formulation recognized as a health functional food.
또한, 상기 목적을 달성하기 위한 본 발명의 또 다른 하나의 양태는 바이오컨버전된 유청 발효물을 유효성분으로 포함하는, 식품 첨가제 조성물을 제공한다. 구체적으로, 상기 유청 발효물은 엔테로코커스 패칼리스 균주로 바이오컨버전된 것일 수 있으며, 보다 구체적으로 수탁번호 KACC 81148BP의 엔테로코커스 패칼리스 M157균주로 바이오컨버전된 것일 수 있다. 상기 용어 엔테로코커스 패칼리스 M157 균주, 바이오컨버전, 유청 및 발효물은 전술한 바와 같다.In addition, another aspect of the present invention for achieving the above object provides a food additive composition comprising bioconverted whey fermentation product as an active ingredient. Specifically, the whey fermentation product may be bioconverted into an Enterococcus faecalis strain, and more specifically, may be bioconverted into an Enterococcus faecalis M157 strain with accession number KACC 81148BP. The terms Enterococcus faecalis M157 strain, bioconversion, whey and fermentation are as described above.
본 발명에서 용어 "식품 첨가제"는 식품을 제조, 가공 또는 보존함에 있어 보존성의 향상, 품질 향상, 영양 강화, 풍미 및 외관을 좋게 하는 등의 목적으로 식품에 첨가, 혼합, 침윤 기타의 방법으로 사용되는 물질을 의미할 수 있다. 상기 식품 첨가제는 본 발명의 유효성분을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있으며, 유효성분의 혼합량은 사용 목적에 따라 통상의 기술자에 의해 적절히 결정될 수 있다. 본 발명의 엔테로코커스 패칼리스 M157 균주로 바이오컨버전된 유청 발효물을 포함하는 식품 첨가제 조성물은 첨가되는 식품 또는 식품 성분에 더하여 구강질환 또는 대장 질환의 예방, 개선 효과를 달성할 수 있다. 본 발명의 또 다른 하나의 양태로서, 본 발명은 유청 발효 시 구강질환 또는 대장질환 개선 효능을 나타내는 수탁번호 KACC 81148BP의 엔테로코커스 패칼리스 M157 균주를 제공한다.In the present invention, the term "food additive" is used by adding, mixing, infiltrating and other methods to food for the purpose of improving preservation, improving quality, enhancing nutrition, improving flavor and appearance when manufacturing, processing or preserving food. It can mean a substance that becomes The food additive can be added as is with the active ingredient of the present invention or used together with other food or food ingredients, and the mixing amount of the active ingredient can be appropriately determined by a person skilled in the art depending on the purpose of use. The food additive composition containing fermented whey bioconverted with Enterococcus faecalis M157 strain of the present invention can achieve the effect of preventing and improving oral diseases or colonic diseases in addition to the added food or food ingredient. In another aspect of the present invention, the present invention provides an Enterococcus faecalis M157 strain with accession number KACC 81148BP, which shows efficacy in improving oral disease or colon disease during whey fermentation.
본 발명에서 엔테로코커스 패칼리스 M157 균주는 전술한 바와 같다.In the present invention, the Enterococcus faecalis M157 strain is as described above.
본 발명의 또 다른 하나의 양태로서, 본 발명은 엔테로코커스 패칼리스 균주로 바이오컨버전된 유청 발효물을 유효성분으로 포함하는, 구강질환 또는 대장질환 예방 또는 치료용 약학적 조성물을 제공한다.In another aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating oral disease or colon disease, comprising fermented whey bioconverted to Enterococcus faecalis strain as an active ingredient.
상기 용어 엔테로코커스 패칼리스 균주, 바이오컨버전, 유청, 발효물, 구강질환, 대장질환 및 예방은 전술한 바와 같다.The terms Enterococcus faecalis strain, bioconversion, whey, fermented product, oral disease, colon disease, and prevention are the same as described above.
본 발명의 약학적 조성물은 엔테로코커스 패칼리스 균주로 바이오컨버전된 유청 발효물 이외에 약학적 조성물의 제조에 통상적으로 사용하는 약학적으로 허용 가능한 담체, 부형제 또는 희석제를 추가로 포함할 수 있다. 상기 담체, 부형제 및 희석제의 구체적인 예로는 락토오스, 덱스트로즈, 수크로오스, 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 또는 광물유 등이 사용될 수 있으며, 이에 특별히 제한되지 않는다. 또한, 본 발명의 약학적 조성물은 각각 통상의 방법에 따라 정제, 환제, 산제, 과립제, 캡슐제, 현탁제, 내용액제, 유제, 시럽제, 멸균된 수용액, 비수성용제 및 동결 건조제로 이루어진 군으로부터 선택되는 어느 하나의 제형을 가질 수 있으며, 상기 제형은 경구 또는 비경구의 여러 가지 형태일 수 있다. 제제화할 경우에는 통상적으로 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제될 수 있으며, 이에 제한되지 않는다. 구체적으로, 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 사용될 수 있으며, 상기 고형제제는 적어도 하나 이상의 부형제 예컨대 전분, 탄산칼슘, 수크로오스 또는 락토오스, 젤라틴 등이 사용될 수 있다. 또한, 단순한 부형제 이외에 스테아린산 마그네슘, 탈크 등과 같은 윤활제 등이 사용될 수 있으며, 이에 제한되지 않는다. 또한, 경구투여를 위한 액상제제로는 현탁제, 내용액제, 유제, 시럽제 등이 사용될 수 있으며, 통상적으로 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예컨대 습윤제, 감미제, 방향제, 보존제 등이 사용될 수 있다. 비경구투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제 또는 좌제 등이 사용될 수 있다. 상기 비수성용제, 현탁제로는 프로필렌글리콜, 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있으며, 이에 제한되지 않는다.The pharmaceutical composition of the present invention may further include pharmaceutically acceptable carriers, excipients, or diluents commonly used in the production of pharmaceutical compositions in addition to the whey fermentation product bioconverted with Enterococcus faecalis strains. Specific examples of the carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline. Cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil may be used, but are not particularly limited thereto. In addition, the pharmaceutical composition of the present invention is selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, solutions for internal use, emulsions, syrups, sterilized aqueous solutions, non-aqueous solvents, and freeze-dried agents, respectively, according to conventional methods. It may have any one dosage form, and the dosage form may be in various oral or parenteral forms. When formulated, it may be prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants, but is not limited thereto. Specifically, solid preparations for oral administration may include tablets, pills, powders, granules, capsules, etc., and the solid preparation may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. . Additionally, in addition to simple excipients, lubricants such as magnesium stearate, talc, etc. may be used, but are not limited thereto. In addition, liquid preparations for oral administration include suspensions, oral solutions, emulsions, syrups, etc., and in addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. This can be used. Preparations for parenteral administration may include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, or suppositories. The non-aqueous solvent or suspension may include, but is not limited to, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate.
본 발명 약학적 조성물의 개별적인 투약의 최적량 및 투약 간격은 구강질환 또는 치주질환의 성질 및 정도, 투여 제형, 경로 및 부위, 그리고 적용 대상의 나이와 건강상태에 의해 결정될 것이고, 의사가 궁극적으로 사용될 적절한 투약을 결정할 것이라는 것은 당해 분야의 당업자가 알 수 있을 것이다. 이러한 투약은 적절할 정도로 자주 반복될 수 있고, 보통의 임상 진료에 따라서 투여량 및 빈도를 변경하거나 또는 감소시킬 수 있다. 또한, 본 발명 조성물의 투여 경로는 본 발명의 목적을 달성할 수 있는 한 어떠한 일반적인 경로를 통하여 투여될 수 있다. 예컨대 본 발명의 약학적 조성물은 복강내 투여, 정맥내 투여, 피하 투여, 피내 투여, 경구 투여될 수 있으나, 이에 제한되지는 않는다.The optimal amount and dosing interval for individual administration of the pharmaceutical composition of the present invention will be determined by the nature and degree of oral disease or periodontal disease, the dosage form, route and site of administration, and the age and health status of the subject, and will be determined by the doctor ultimately. It will be appreciated by those skilled in the art to determine appropriate dosage. Such administration may be repeated as often as appropriate, and the dosage and frequency may be varied or reduced according to normal clinical practice. Additionally, the administration route of the composition of the present invention may be administered through any general route as long as the purpose of the present invention can be achieved. For example, the pharmaceutical composition of the present invention may be administered intraperitoneally, intravenously, subcutaneously, intradermally, or orally, but is not limited thereto.
본 발명에서 용어 "치료"는 본 발명 조성물의 투여로 구강질환 또는 대장질환의 증상이 호전되거나 이롭게 변경되는 모든 행위를 의미한다.In the present invention, the term “treatment” refers to any action in which the symptoms of oral disease or colon disease are improved or beneficially changed by administration of the composition of the present invention.
본 발명의 또 다른 하나의 양태로서, 본 발명은 엔테로코커스 패칼리스 균주로 바이오컨버전된 유청 발효물을 유효성분으로 포함하는, 구강질환 또는 대장질환 예방 또는 개선용 의약외품 조성물을 제공한다.In another aspect of the present invention, the present invention provides a quasi-drug composition for preventing or improving oral disease or colon disease, comprising fermented whey bioconverted to Enterococcus faecalis strain as an active ingredient.
본 발명에 있어서, 상기 의약외품의 종류는 특별히 제한되지 아니하며, 당해 기술 분야에서 통상적으로 사용되는 의약외품일 수 있다. 상기 의약외품의 비제한적인 예로는, 치약, 구강청결제, 껌, 캔디류, 구강스프레이, 구강용 연고제, 구강용 바니쉬(varnish), 구강필름 및 잇몸 마사지 크림으로 구성된 군으로부터 선택되는 하나 이상의 제형일 수 있다. 이들은 단독으로 사용되거나 2종 이상을 혼합하여 사용될 수 있다.In the present invention, the type of the quasi-drug is not particularly limited, and may be a quasi-drug commonly used in the relevant technical field. Non-limiting examples of the quasi-drugs may be one or more formulations selected from the group consisting of toothpaste, mouthwash, gum, candy, oral spray, oral ointment, oral varnish, oral film, and gum massage cream. . These may be used alone or in combination of two or more types.
본 발명의 또 다른 하나의 양태로서, 본 발명은 엔테로코커스 패칼리스 균주로 바이오컨버전된 유청 발효물을 유효성분으로 포함하는, 구강질환 또는 대장질환 예방 또는 개선용 사료 조성물을 제공한다.In another aspect of the present invention, the present invention provides a feed composition for preventing or improving oral disease or colonic disease, comprising fermented whey bioconverted to Enterococcus faecalis strain as an active ingredient.
상기 사료 조성물은 사료 첨가제를 포함할 수 있다. 본 발명의 사료첨가제는 사료관리법상의 보조사료에 해당한다.The feed composition may include feed additives. The feed additive of the present invention corresponds to supplementary feed under the Feed Management Act.
본 발명에서 용어, "사료"는 특히 동물이 먹고, 섭취하며, 소화시키기 위한 또는 이에 적당한 임의의 천연 또는 인공 규정식, 한끼식 등 또는 상기 한끼식의 성분을 의미할 수 있다.In the present invention, the term "feed" may mean any natural or artificial diet, meal, etc., or a component of the meal, especially for or suitable for eating, ingestion, and digestion by animals.
상기 사료의 종류는 특별히 제한되지 아니하며, 당해 기술 분야에서 통상적으로 사용되는 사료를 사용할 수 있다. 상기 사료의 비제한적인 예로는, 곡물류, 근과류, 식품 가공 부산물류, 조류, 섬유질류, 제약 부산물류, 유지류, 전분류, 박류 또는 곡물 부산물류 등과 같은 식물성 사료; 단백질류, 무기물류, 유지류, 광물성류, 유지류, 단세포 단백질류, 동물성 플랑크톤류 또는 음식물 등과 같은 동물성 사료를 들 수 있다. 이들은 단독으로 사용되거나 2 종 이상을 혼합하여 사용될 수 있다.The type of feed is not particularly limited, and feed commonly used in the art can be used. Non-limiting examples of the feed include plant feeds such as grains, roots and fruits, food processing by-products, algae, fiber, pharmaceutical by-products, oils and fats, starches, cucurbits or grain by-products; Examples include animal feeds such as proteins, inorganic substances, fats and oils, minerals, oils and fats, single-cell proteins, zooplanktons or food. These may be used individually or in combination of two or more types.
본 발명에서 엔테로코커스 패칼리스 M157 균주로 바이오컨버전된 유청 발효물은 치주조직의 총 부피 당 골 부피 증가, 골 부피 당 표면적 감소, 골 소실 수피 감소 효과를 나타내며 치주 조직 및 대장 조직에서 전염증 지표를 감소시키고 항염증 지표를 증가시키는 바, 구강 질환 및 대장 질환의 예방, 개선 또는 치료용으로 유용하게 사용될 수 있어 산업적 활용 가치가 높다.In the present invention, the whey fermentation product bioconverted to Enterococcus faecalis M157 strain increases bone volume per total volume of periodontal tissue, decreases surface area per bone volume, reduces bone loss and bark, and exhibits pro-inflammatory indicators in periodontal tissue and colon tissue. As it reduces and increases anti-inflammatory indicators, it can be useful for preventing, improving or treating oral diseases and colon diseases, so it has high industrial value.
도 1은 정상군, 대조군 및 M157군에 대한 치주 조직의 Micro-CT 분석 결과를 나타낸 것이다.
도 2는 정상군, 대조군 및 M157군에 대한 치주 조직의 광학 현미경 사진을 나타낸 것이다.
도 3은 정상군, 대조군 및 M157군에 대한 치주 조직에서의 면역 지표 전사체 발현량을 분석한 결과를 나타낸 것이다.
도 4는 정상군, 대조군 및 M157군에 대한 대장 조직에서의 면역 지표 전사체 발현량을 분석한 결과를 나타낸 것이다.
도 5는 정상군, 대조군 및 M157군에 대한 치주 조직에서의 면역 지표 단백질 발현량을 분석한 결과를 나타낸 것이다.
도 6는 정상군, 대조군 및 M157군에 대한 대장 조직에서의 면역 지표 단백질 발현량을 분석한 결과를 나타낸 것이다. Figure 1 shows the results of Micro-CT analysis of periodontal tissue for the normal group, control group, and M157 group.
Figure 2 shows optical micrographs of periodontal tissue for the normal group, control group, and M157 group.
Figure 3 shows the results of analyzing the expression levels of immune indicator transcripts in periodontal tissues for the normal group, control group, and M157 group.
Figure 4 shows the results of analyzing the expression levels of immune indicator transcripts in colonic tissue for the normal group, control group, and M157 group.
Figure 5 shows the results of analyzing the expression level of immune indicator proteins in periodontal tissue for the normal group, control group, and M157 group.
Figure 6 shows the results of analyzing the expression levels of immune indicator proteins in colonic tissue for the normal group, control group, and M157 group.
이하, 본 발명을 하기 실시예를 통하여 보다 상세하게 설명한다. 그러나 이들 실시예는 본 발명을 예시적으로 설명하기 위한 것으로 본 발명의 범위가 이들 실시예만으로 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail through the following examples. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.
실시예 1: 엔테로코커스 패칼리스 M157 균주를 활용한 유청발효 산물 제조Example 1: Preparation of whey fermentation product using Enterococcus faecalis M157 strain
분리균주 엔테로코커스 패칼리스 M157을 이용하여 유청발효 산물(이하 바이오컨버전 산물)을 제조하였다. 10 mL의 MRS broth에서 37℃, 24시간 호기 배양한 뒤, 유청 분말을 이용해 제조 후 살균한 10% whey 용액에 1%의 농도가 되도록 접종하였다. 37℃, 24시간 배양한 뒤, 1N NaOH (Duksan general science, Seoul, Korea)를 이용하여 pH 6.5로 조정한 후 30%로 농축하여 분무건조기로 분말화하여 바이오컨버전 산물로 사용하였다. 바이오컨버전 산물을 1,500 mg/10 mL/kg의 농도가 되도록 phosphate buffered saline (PBS)로 현탁하여 경구 투여 및 치아 도포 시료로 사용하였다.Whey fermentation products (hereinafter referred to as bioconversion products) were prepared using the isolated strain Enterococcus faecalis M157. After aerobically cultivating in 10 mL of MRS broth at 37°C for 24 hours, it was prepared using whey powder and then inoculated into a sterilized 10% whey solution at a concentration of 1%. After culturing at 37°C for 24 hours, the pH was adjusted to 6.5 using 1N NaOH (Duksan general science, Seoul, Korea), concentrated to 30%, powdered with a spray dryer, and used as a bioconversion product. The bioconversion product was suspended in phosphate buffered saline (PBS) to a concentration of 1,500 mg/10 mL/kg and used as a sample for oral administration and dental application.
실시예 2: 유청 바이오컨버전 산물에 의한 구강질환 및 대장질환 개선 효과 확인Example 2: Confirmation of improvement effect of oral disease and colon disease by whey bioconversion product
2-1. 치주염 동물모델 제조2-1. Manufacturing of periodontitis animal model
1주간 적응기간과 2일간 항생제 식수 제공 기간 후 7 주령의 랫드에 zoletile (1 mg/mL) (Virbac, France)과 rompun (Bayer, Germany)[4:1(v/v)]을 1 mL/kg으로 복강 내 주사하여 마취시킨 후 우측 하악 제 1 대구치에 멸균된 봉합사로 결찰하여 치주염을 유발하였다. 구체적인 실험 모델은 표 1에서 나타낸 바와 같다.After a 1-week adaptation period and a 2-day antibiotic drinking period, 7-week-old rats were administered zoletile (1 mg/mL) (Virbac, France) and rompun (Bayer, Germany) [4:1(v/v)] at 1 mL/mL. After anesthetizing the patient by intraperitoneal injection (kg), periodontitis was induced by ligating the right mandibular first molar with sterilized suture. The specific experimental model is shown in Table 1.
M157 유청발효산물Enterococcus faecalis
M157 Whey fermentation product
2-2. Micro-CT 분석에 의한 치주염 개선 효과 확인2-2. Confirmation of periodontitis improvement effect through Micro-CT analysis
바이오컨버전 산물의 치주염 개선 효과를 확인하기 위하여, 치주 조직에서 정상군, 대조군 및 M157 유청발효산물 처리군 (이하, M157군) 각각 3 마리를 이용하여 micro-CT 분석을 실시하였다. Micro-CT 분석을 위해 치주 조직을 10% 중성 formalin으로 고정시킨 후 치조골 소실 부피를 측정하기 위해 Quantum FX microCT scanner(Perkin-Elmer, Waltham, MA, USA)를 이용하여 2차원 평면 영상을 측정하였고 CTAn software(Skyscan, AaRTselaar, Belgium)를 사용하여 치주 조직의 총 부피 당 골 부피(BV/TV), 골 부피 당 표면적(BS/BV), 골 소실 부피 등을 분석하였다.In order to confirm the effect of the bioconversion product on improving periodontitis, micro-CT analysis was performed on periodontal tissue using three animals each: the normal group, the control group, and the M157 whey fermentation product treatment group (hereinafter referred to as the M157 group). For Micro-CT analysis, periodontal tissue was fixed with 10% neutral formalin, and then two-dimensional planar images were measured using a Quantum FX microCT scanner (Perkin-Elmer, Waltham, MA, USA) to measure the volume of alveolar bone loss, and CTAn Bone volume per total volume of periodontal tissue (BV/TV), surface area per bone volume (BS/BV), and bone loss volume were analyzed using software (Skyscan, AaRTselaar, Belgium).
Micro-CT 분석 결과, 표 2 및 도 1에 나타낸 바와 같이, 피질골의 양과 비례하는 총 부피 당 골 부피(BV/TV)가 대조군 대비 정상군, M157군에서 유의적으로 높게 측정되었다(p<0.05). 소주골의 양과 비례하는 골 부피 당 표면적(BS/BV)은 대조군 대비 정상군, M157군에서 유의적으로 낮게 측정되었다(p<0.05). 골 소실 부피(Bone loss volume) 분석 결과, 대조군 대비 정상군 및 M157군에서 유의적으로 낮게 측정되었다(p<0.05). As a result of Micro-CT analysis, as shown in Table 2 and Figure 1, bone volume per total volume (BV/TV), which is proportional to the amount of cortical bone, was measured to be significantly higher in the normal group and M157 group compared to the control group (p<0.05 ). The surface area per bone volume (BS/BV), which is proportional to the amount of trabecular bone, was measured to be significantly lower in the normal and M157 groups compared to the control group (p<0.05). As a result of bone loss volume analysis, it was measured to be significantly lower in the normal group and M157 group compared to the control group (p<0.05).
결과적으로, M157군에서 BS/TV, BS/BV, Bone loss volume이 정상군과 유의적 차이가 없으며 대조군보다 개선된 상태를 나타내어, E. faecalis M157로 제조한 바이오컨버전 산물이 치주염 개선에 효과가 있는 것으로 확인되었다.As a result, in the M157 group, BS/TV, BS/BV, and bone loss volume were not significantly different from the normal group and showed improvement compared to the control group, indicating that the bioconversion product manufactured with E. faecalis M157 was effective in improving periodontitis. It was confirmed that there was.
(BS/BV) (1/mm)(BS/BV) (1/mm)
(mm(mm
33
))
2-3. 조직병리학적 분석에 의한 치주염 개선 효과 확인2-3. Confirmation of periodontitis improvement effect through histopathological analysis
바이오컨버전 산물 투여와 도포에 따른 치주염 개선 효과를 검증하기 위해 치주염 유발 랫드의 치주 조직에서 조직형태학적 변화, 치조골, 파골세포, 조골세포 수 등을 측정하였다. To verify the effect of improving periodontitis by administering and applying the bioconversion product, histomorphological changes, alveolar bone, osteoclasts, and osteoblast numbers were measured in the periodontal tissue of periodontitis-induced rats.
구체적으로, 치주염으로 인한 염증 발생 유무 확인을 위해 Mecklenburg et al.(2013)을 참고하여 조직병리학적 분석을 실시하였다. 삭정, 탈수, 파라핀 포매, 박절 등 일반적인 조직처리과정을 거쳐 제작한 검체에 H&E staining 후 광학현미경(Olympus BX53, Japan)을 이용하여 조직병리학적 변화를 관찰하였다. Specifically, histopathological analysis was performed referring to Mecklenburg et al. (2013) to confirm whether inflammation occurred due to periodontitis. Histopathological changes were observed using an optical microscope (Olympus BX53, Japan) after H&E staining of specimens prepared through general tissue processing procedures such as trimming, dehydration, paraffin embedding, and sectioning.
그 결과, 표 3 및 도 2에 나타낸 바와 같이, 대조군에서 염증세포 침윤과 상피증식이 가장 많이 관찰되었으며, M157군에서 정상군과 마찬가지로 병변이 확인되지 않아 E. faecalis M157로 제조한 바이오컨버전 산물의 우수한 치주염 개선 효과를 확인하였다.As a result, as shown in Table 3 and Figure 2, the most inflammatory cell infiltration and epithelial proliferation were observed in the control group, and no lesions were identified in the M157 group like the normal group, so that the bioconversion product manufactured with E. faecalis M157 Excellent periodontitis improvement effect was confirmed.
2-4. 전사체 분석에 의한 구강질환 및 대장질환 개선 효과 확인2-4. Confirmation of improvement in oral and colonic diseases through transcriptome analysis
바이오컨버전 산물 투여로 인한 치주 조직 및 대장 조직에서의 염증 및 산화적 스트레스 개선을 확인하기 위해 전사체 분석을 실시하였다. Transcriptome analysis was conducted to confirm the improvement of inflammation and oxidative stress in periodontal tissue and colonic tissue due to administration of the bioconversion product.
구체적으로, 치주 조직과 대장 조직에서 RNeasy mini kit(Qiagen)을 이용하여 mRNA를 추출하였고 Quantitect Reverse Transcription kit(Qiagen)을 이용하여 cDNA를 합성한 후 전사체 분석에 이용하였다. cDNA로 RT-PCR을 진행하여 Ct 수치를 구하였고 2 -ΔΔCt 방법으로 전사체 발현량을 계산하였다. Specifically, mRNA was extracted from periodontal tissue and colonic tissue using the RNeasy mini kit (Qiagen), and cDNA was synthesized using the Quantitect Reverse Transcription kit (Qiagen) and used for transcriptome analysis. RT-PCR was performed with cDNA to obtain Ct levels, and transcript expression levels were calculated using the 2 -ΔΔCt method.
그 결과, 도 3에 나타낸 바와 같이, 치주조직에서 전염증 지표 TNF-α, IL-6 및 IL-1β의 전사체 발현량은 대조군 대비 M157군에서 유의적으로 낮게 관찰되었다(p<0.05).As a result, as shown in Figure 3, the transcript expression levels of pro-inflammatory indicators TNF-α, IL-6, and IL-1β in periodontal tissue were observed to be significantly lower in the M157 group compared to the control group (p<0.05).
또한, 도 4에 나타낸 바와 같이, 대장조직에서 전염증 지표 TNF-α 및 IL-1β의 전사체 발현량은 대조군 대비 M157군에서 낮게 관찰되었다(p<0.05).In addition, as shown in Figure 4, the transcript expression levels of pro-inflammatory indicators TNF-α and IL-1β in colon tissue were observed to be lower in the M157 group compared to the control group (p<0.05).
이러한 결과에 따라 E. faecalis M157로 제조한 바이오컨버전 산물은 치주 및 대장 조직에서 염증 억제 효과가 있는 것을 확인하였다.According to these results, it was confirmed that the bioconversion product prepared from E. faecalis M157 has an anti-inflammatory effect in periodontal and colonic tissues.
2-5. 단백질 분석에 의한 구강질환 및 대장질환 개선 효과 확인2-5. Confirmation of improvement in oral and colonic diseases through protein analysis
바이오컨버전 산물 투여로 인한 치주 조직 및 대장 조직의 염증 및 산화적 스트레스의 개선을 확인하기 위해 단백질 발현량을 이용하여 면역 지표(TNF-α, IL-6, IL-10, SOD1, GPx1)를 분석하였다.To confirm the improvement of inflammation and oxidative stress in periodontal and colonic tissues due to administration of bioconversion products, immune indicators (TNF-α, IL-6, IL-10, SOD1, GPx1) were analyzed using protein expression levels. did.
치주 조직 단백질에서의 면역 지표 분석 결과, 도 5에 나타낸 바와 같이,As a result of immune indicator analysis in periodontal tissue proteins, as shown in Figure 5,
M157군에서 전염증 지표 TNF-α 및 IL-6는 대조군 대비 유의적으로 낮게 관찰되었고, 정상군 대비 유의적인 차이를 나타내지 않았다(p<0.05). 또한 항산화 지표 GPx1 분석 결과, 대조군 대비 M157군에서 높은 발현량이 확인되었다(p<0.05).In the M157 group, pro-inflammatory indicators TNF-α and IL-6 were observed to be significantly lower than the control group, and showed no significant difference compared to the normal group (p<0.05). In addition, as a result of analysis of the antioxidant index GPx1, higher expression level was confirmed in the M157 group compared to the control group (p<0.05).
대장 조직 단백질에서의 면역 지표 분석 결과, 도 6에 나타낸 바와 같이. M157군에서 전염증 지표 TNF-α는 대조군 대비 유의적으로 낮은 발현량이 확인되었다(p<0.05). 항염증 지표 IL-10 및 SOD1의 경우, 대조군 대비 M157군에서 유의적으로 높은 발현량이 확인되었다(p<0.05). Results of immune indicator analysis in colon tissue proteins, as shown in Figure 6. In the M157 group, the pro-inflammatory marker TNF-α was found to have a significantly lower expression level compared to the control group (p<0.05). In the case of anti-inflammatory indicators IL-10 and SOD1, significantly higher expression levels were confirmed in the M157 group compared to the control group (p<0.05).
이러한 결과를 통해 E. faecalis M157 균주로 제조한 바이오컨버전 산물이 치주 조직에서 염증 억제와 산화적 스트레스에 효과가 있으며, 대장 조직의 염증 개선 및 예방 효과가 있음을 확인하였다. Through these results, it was confirmed that the bioconversion product manufactured with E. faecalis M157 strain is effective in suppressing inflammation and oxidative stress in periodontal tissue, and has an effect in improving and preventing inflammation in colon tissue.
이상의 설명으로부터, 본 발명이 속하는 기술분야의 당업자는 본 발명이 그 기술적 사상이나 필수적 특징을 변경하지 않고서 다른 구체적인 형태로 실시될 수 있다는 것을 이해할 수 있을 것이다. 이와 관련하여, 이상에서 기술한 실시예들은 모든 면에서 예시적인 것이며 한정적인 것이 아닌 것으로 이해해야만 한다. 본 발명의 범위는 상기 상세한 설명보다는 후술하는 특허 청구범위의 의미 및 범위 그리고 그 등가 개념으로부터 도출되는 모든 변경 또는 변형된 형태가 본 발명의 범위에 포함되는 것으로 해석되어야 한다.From the above description, those skilled in the art to which the present invention pertains will understand that the present invention can be implemented in other specific forms without changing its technical idea or essential features. In this regard, the embodiments described above should be understood in all respects as illustrative and not restrictive. The scope of the present invention should be construed as including the meaning and scope of the patent claims described below rather than the detailed description above, and all changes or modified forms derived from the equivalent concept thereof are included in the scope of the present invention.
Claims (14)
상기 구강질환은 치조골 파손(alveolar bone breakage), 치조골다공증(alveolar bone osteoporosis), 치조골연화증(alveolar bone osteomalacia), 치조골감소증(alveolar bone osteopenia), 치아 우식증(dental caries), 치은염(gingivitis) 및 치주염(periodontitis)으로 구성된 군으로부터 선택되는 하나 이상의 질환인 것인, 식품 조성물.
A food composition for preventing or improving oral diseases, comprising as an active ingredient whey fermentation product bioconverted with Enterococcus faecalis M157 strain (accession number KACC 81148BP),
The oral diseases include alveolar bone breakage, alveolar bone osteoporosis, alveolar bone osteomalacia, alveolar bone osteopenia, dental caries, gingivitis and periodontitis ( A food composition, which is at least one disease selected from the group consisting of periodontitis.
상기 구강질환은 치조골 파손(alveolar bone breakage), 치조골다공증(alveolar bone osteoporosis), 치조골연화증(alveolar bone osteomalacia), 치조골감소증(alveolar bone osteopenia), 치아 우식증(dental caries), 치은염(gingivitis) 및 치주염(periodontitis)으로 구성된 군으로부터 선택되는 하나 이상의 질환인 것인, 건강기능식품.
A health functional food for preventing or improving oral diseases, containing as an active ingredient whey fermentation product bioconverted to Enterococcus faecalis M157 strain (accession number KACC 81148BP),
The oral diseases include alveolar bone breakage, alveolar bone osteoporosis, alveolar bone osteomalacia, alveolar bone osteopenia, dental caries, gingivitis and periodontitis ( A health functional food that is one or more diseases selected from the group consisting of periodontitis.
상기 건강기능식품은 정제, 캡슐, 환, 과립, 액상, 분말, 편상, 페이스트상, 시럽, 겔, 젤리, 바 등의 건강기능식품 제제류, 음료류, 껌류 및 캔디류로 구성된 군으로부터 선택되는 하나 이상의 제형인 것인, 건강기능식품.
According to paragraph 4,
The health functional food is one or more selected from the group consisting of health functional food preparations such as tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, bars, beverages, gums, and candies. A health functional food in dosage form.
상기 구강질환은 치조골 파손(alveolar bone breakage), 치조골다공증(alveolar bone osteoporosis), 치조골연화증(alveolar bone osteomalacia), 치조골감소증(alveolar bone osteopenia), 치아 우식증(dental caries), 치은염(gingivitis) 및 치주염(periodontitis)으로 구성된 군으로부터 선택되는 하나 이상의 질환인 것인, 식품 첨가제 조성물.
A food additive composition for preventing or improving oral diseases, comprising fermented whey bioconverted with Enterococcus faecalis M157 strain (accession number KACC 81148BP) as an active ingredient,
The oral diseases include alveolar bone breakage, alveolar bone osteoporosis, alveolar bone osteomalacia, alveolar bone osteopenia, dental caries, gingivitis and periodontitis ( A food additive composition, which is one or more diseases selected from the group consisting of periodontitis.
상기 구강질환은 치조골 파손(alveolar bone breakage), 치조골다공증(alveolar bone osteoporosis), 치조골연화증(alveolar bone osteomalacia), 치조골감소증(alveolar bone osteopenia), 치아 우식증(dental caries), 치은염(gingivitis) 및 치주염(periodontitis)으로 구성된 군으로부터 선택되는 하나 이상의 질환인 것인, 약학적 조성물.
A pharmaceutical composition for preventing or treating oral diseases, comprising fermented whey bioconverted with Enterococcus faecalis M157 strain (accession number KACC 81148BP) as an active ingredient,
The oral diseases include alveolar bone breakage, alveolar bone osteoporosis, alveolar bone osteomalacia, alveolar bone osteopenia, dental caries, gingivitis and periodontitis ( A pharmaceutical composition, which is one or more diseases selected from the group consisting of periodontitis.
상기 구강질환은 치조골 파손(alveolar bone breakage), 치조골다공증(alveolar bone osteoporosis), 치조골연화증(alveolar bone osteomalacia), 치조골감소증(alveolar bone osteopenia), 치아 우식증(dental caries), 치은염(gingivitis) 및 치주염(periodontitis)으로 구성된 군으로부터 선택되는 하나 이상의 질환인 것인, 의약외품 조성물.
A quasi-drug composition for preventing or improving oral diseases, comprising fermented whey bioconverted with Enterococcus faecalis M157 strain (accession number KACC 81148BP) as an active ingredient,
The oral diseases include alveolar bone breakage, alveolar bone osteoporosis, alveolar bone osteomalacia, alveolar bone osteopenia, dental caries, gingivitis and periodontitis ( A quasi-drug composition, which is one or more diseases selected from the group consisting of periodontitis.
상기 의약외품 조성물은 치약, 구강청결제, 껌, 캔디류, 구강스프레이, 구강용 연고제, 구강용 바니쉬(varnish), 구강필름 및 잇몸 마사지 크림으로 구성된 군으로부터 선택되는 하나 이상의 제형인 것인, 의약외품 조성물.
According to clause 12,
The quasi-drug composition is one or more formulations selected from the group consisting of toothpaste, mouthwash, gum, candy, oral spray, oral ointment, oral varnish, oral film, and gum massage cream. A quasi-drug composition.
상기 구강질환은 치조골 파손(alveolar bone breakage), 치조골다공증(alveolar bone osteoporosis), 치조골연화증(alveolar bone osteomalacia), 치조골감소증(alveolar bone osteopenia), 치아 우식증(dental caries), 치은염(gingivitis) 및 치주염(periodontitis)으로 구성된 군으로부터 선택되는 하나 이상의 질환인 것인, 사료 조성물.A feed composition for preventing or improving oral diseases, comprising fermented whey bioconverted with Enterococcus faecalis M157 strain (accession number KACC 81148BP) as an active ingredient,
The oral diseases include alveolar bone breakage, alveolar bone osteoporosis, alveolar bone osteomalacia, alveolar bone osteopenia, dental caries, gingivitis and periodontitis ( A feed composition, which is one or more diseases selected from the group consisting of periodontitis.
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