KR20160140471A - A bioconversion product comprising Tumeric - Google Patents
A bioconversion product comprising Tumeric Download PDFInfo
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- KR20160140471A KR20160140471A KR1020160065076A KR20160065076A KR20160140471A KR 20160140471 A KR20160140471 A KR 20160140471A KR 1020160065076 A KR1020160065076 A KR 1020160065076A KR 20160065076 A KR20160065076 A KR 20160065076A KR 20160140471 A KR20160140471 A KR 20160140471A
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- enzyme
- salmonella
- rice bran
- mushroom
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Abstract
Description
본 발명은 미생물(담자균류균사) 발효 및 효소처리 생물전환공정을 통해 생물전환된 미강생물전환산물에 관한 것이다.
The present invention relates to a microorganism (microbial mycelium) fermentation and an organism bioconversion product bioconverted through an enzyme-treated biological conversion process.
축산물은 사람의 주요 식품원으로서 중요하지만, 식중독균의 주요 매개체이기도 하다. 살모넬라 감염증은 미국과 유럽에서는 수인성 식품매개질환 중 가장 높은 비율을 차지하고 있으며, 2008년도 국내 조사 결과에서는 두 번째로 높은 비율을 차지할 정도로 공중보건학적으로 많은 문제를 야기하고 있다. FAO/WHO(2002) 보고에 따르면 유럽에서 발생하는 식중독의 약 26%가 계란 및 계육을 포함한 가금 유래이며, 식중독 발생의 77.1%가 살모넬라 균이 원인으로 나타났다. 주요 살모넬라 원인균은 S. Enteritidis로, 1/3 이상을 차지하는 것으로 나타났으며, 미국에서는 다른 식중독 유발 세균에 비해 치사율이 높은 원인으로 파악되고 있다. 국내에서도 닭도체로부터 살모넬라 분석 결과, 분리한 24종 중 S. Enteritidis가 17종(70.8%)으로 가장 높게 나타났다. 살모넬라는 가금류뿐만 아니라, 육류를 통해서도 문제를 일으키고 있는데 2008년 유럽의 보고서에 의하면 살모넬라증 사례는 원인 별로는 대략 가금류에 의한 살모넬라 오염이 50~70%를 차지하고 있으며, 그 뒤를 돼지고기와 소고기가 각각 10~20%씩 같은 비율을 차지하고 있다. Livestock products are important as a major source of food for humans, but they are also a major mediator of food poisoning bacteria. Salmonella infections account for the highest percentage of water-borne foodborne illnesses in the United States and Europe, and are the second highest percentage in domestic surveys in 2008, causing many public health problems. According to the FAO / WHO (2002) report, about 26% of food poisoning in Europe is from poultry, including eggs and meat, and 77.1% of food poisoning cases are due to Salmonella. S. enteritidis was the major cause of Salmonella, accounting for more than one - third of the cases. In the United States, the cause of death is higher than other foodborne pathogens. In Korea, S. enteritidis was the highest among 17 isolates (70.8%) among 24 isolates from chicken carcasses. Salmonella is causing problems not only by poultry but also by meat. According to a European report in 2008, salmonella cases accounted for about 50 ~ 70% of the cases of salmonella contamination by poultry, followed by pork and beef by 10 ~ 20%, respectively.
양돈장에서 살모넬라속 균의 감염은, 돼지에게 종 특이성이 있는 살모넬라로 급성 패혈증을 일으켜서 높은 폐사율을 발생시키는 S. Choleraesuis와, 인수공통 살모넬라로 결장염을 일으켜서 설사를 유발하는 S. Typhimurium 및 S. Enteritidis 등에 의해서 주로 일어난다. 특히, 인수공통원인균에 의한 감염은 사람에게는 돈육 오염에 의한 식중독의 원인이 될 수 있어 식육위생 면에서도 중요한 질병이다. S. Choleraesuis는 전세계적으로 임상 증상을 보이는 돼지에서 가장 많이 분리되는 살모넬라임에도 불구하고 국내의 경우 이 균에 의한 질병 발생은 그 빈도가 매우 낮은 것으로 파악되고 있다. Infection of Salmonella spp. In pig farms is a species-specific Salmonella infection in pigs, S. choleraesuis causing acute septicemia resulting in high mortality and S. typhimurium and S. enteritidis causing diarrhea due to colonization with acquired salmonella . In particular, infections caused by common causative microorganisms can cause food poisoning due to pork contamination in humans, which is an important disease in terms of food hygiene. Although S. choleraesuis is the most isolated salmonella in pigs with clinical symptoms worldwide, the incidence of this disease is very low in Korea.
돼지에서 주로 만성 소화기계 질병을 일으키는 S. Typhimurium의 경우는 매우 다양한 오염원이 존재하는데, 오염된 사료나 물 또는 농장 주변의 야생동물(새, 설치류 등)이나 별 증상 없이 균을 보유하고 있는 성돈이 중요한 전염원이 될 수 있다. 특히 건강 보균돈의 감별 진단이 어려울 뿐만 아니라 이균의 숙주가 무한히 많고 저항성이 강하다 보니 도처에 생재할 가능성이 높다.S. Typhimurium, which causes predominantly chronic gastrointestinal disease in pigs, has a wide variety of contaminants, including polluted feed, water, or wild animals around the farm (birds, rodents, etc.) It can be an important infectious source. In particular, it is not only difficult to differentiate the health insurance funds, but also because there are infinite number of hosts and a strong resistance.
세계적으로 세균성 식중독의 발생이 증가하고 있으며, 통계에 따르면 2009년 미국에서 17,468건의 세균성 식중독이 발생하였으며, 살모넬라 식중독은 전체 식중독 발생 건수 중 가장 높은 비율을 차지하고 있다. 또한, 살모넬라 혈청형 분석 결과 6,371개(90.5%) 시료 중 Enteritidis 1,226(19.2%), Typhimurium 1,024(16.1%), Newport 772(12.1%), Javiana 544(8.5%), Heidelberg 230(3.6%), Montevideo 206(3.2%), Muenchen 170(2.7%), Saintpaul 157(2.5%), Oranienburg 132(2.1%)로 나타나, S. Enteritidis 와 S. Typhimurium이 살모넬라에 의한 식중독 발생의 35.2%를 차지하는 것으로 나타났다(CDC, 2010). Bacterial food poisoning has increased worldwide, and statistics show that there were 17,468 bacterial food poisoning cases in the United States in 2009, and Salmonella food poisoning is the highest percentage of all food poisoning cases. Enteritidis 1,226 (19.2%), Typhimurium 1,024 (16.1%), Newport 772 (12.1%), Javiana 544 (8.5%) and Heidelberg 230 (3.6%) were among the 6,371 serotypes of Salmonella serotypes Montevideo 206 (3.2%), Muenchen 170 (2.7%), Saintpaul 157 (2.5%) and Oranienburg 132 (2.1%) while S. Enteritidis and S. Typhimurium accounted for 35.2% of food poisoning cases caused by Salmonella (CDC, 2010).
살모넬라균은 살아있는 닭, 오리, 돼지 및 도체를 통해 도축장 및 가공 처리장으로 유입되어, 작업 라인을 따라 시설을 오염시켜 최종 생산된 축산식품의 미생물적인 품질을 위태롭게 한다. 실제 국내 환경에서도 S. Enteritidis를 비롯한 다양한 인수공통 살모넬라가 분리되고 있으며, 종계장 유래의 살모넬라가 부화장, 육계 사육농장을 거쳐 도축장까지 전달된다는 조사 결과를 고려할 때, 도축장, 농장을 비롯한 생산단계별 통제가 아닌 종계 사육부터 가공장에 이르기까지 모든 단계의 체계적인 통제 기술이 필요하다.Salmonella enters live slaughterhouses and processing plants through live chickens, ducks, pigs, and conductors, which pollute the facility along the line and endanger the microbiological quality of the finished livestock products. Considering the fact that Salmonella from various sources including Salmonella from S. enteritidis has been isolated in the domestic environment and Salmonella derived from the slaughterhouse has been transferred to slaughterhouses through hatcheries and broiler farms, Systematic control techniques at all stages from breeding to processing are needed.
살모넬라의 제어를 위해 백신, 경쟁적 배제제(Competitive exclusion bacteria), 프로바이오틱스, 프리바이오틱스, 베타글루칸, 단쇄지방산(유기산), 중쇄지방산, 에센셜 오일(essential oil), 박테리오신(bacteriocins), 박테리오파지(bacteriophages) 및 항생제 등이 이용되고 있으나(미국 등록특허 US 8062645, 한국 등록특허 KR 542484, 한국 등록특허 KR 941892, 한국 등록특허 KR 1178173), 살모넬라에 의한 식중독이 계속 발생하고 있으므로 이에 대한 새로운 대안이 요구되는 실정이다.Bacteriocins, bacteriophages, antioxidants, antioxidants, antioxidants, antioxidants, antioxidants, antioxidants, antioxidants, antioxidants, antioxidants, antioxidants, And antibiotics have been used (US registered patent US 8062645, Korean registered patent KR 542484, Korean registered patent KR 941892, Korean registered patent KR 1178173), and food poisoning by salmonella continues to occur. to be.
따라서 본 발명은 미생물(담자균류균사) 발효 및 효소처리 생물전환공정을 통해 생물전환된 미강생물전환산물에 관한 것으로, 본 발명의 미강생물전환산물은 살모넬라 감염 및 패혈증 발병 저감에 큰 효과가 있으므로, 안전한 축산물 생산에 크게 기여할 것으로 기대된다.
Accordingly, the present invention relates to a microorganism (microbial mycelium) fermentation product and an enzyme-treated microbial conversion product obtained by biosynthesis process. Since the microbial bioconversion product of the present invention has a great effect on reducing salmonella infection and sepsis, It is expected to contribute greatly to the production of safe livestock products.
본 발명은 상기와 같은 종래의 기술상의 문제점을 해결하기 위해 안출된 것으로, 미생물(담자균류균사) 발효 및 효소처리 생물전환공정을 통해 생물전환된 미강생물전환산물에 관한 것이다. 그러나 본 발명이 이루고자 하는 기술적 과제는 이상에서 언급한 과제에 제한되지 않으며, 언급되지 않은 또 다른 과제들은 아래의 기재로부터 당 업계에서 통상의 지식을 가진 자에게 명확하게 이해될 수 있을 것이다.
Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made to solve the above-mentioned problems occurring in the prior art, and it relates to bioconversion products of microorganisms (mycelium mycelium) fermentation and bioconverted microorganisms through biosynthesis processes. However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.
이하, 본원에 기재된 다양한 구체예가 도면을 참조로 기재된다. 하기 설명에서, 본 발명의 완전한 이해를 위해서 다양한 특이적 상세사항 예컨대, 특이적 형태, 조성물 및 공정 등이 기재되어 있다. 그러나 특정의 구체예는 이들 특이적 상세사항 중 하나 이상 없이, 또는 다른 공지된 방법 및 형태와 함께 실행될 수 있다. 다른 예에서 공지된 공정 및 제조 기술은 본 발명을 불필요하게 모호하게 하지 않게 하기 위해서 특정의 상세사항으로 기재되지 않는다. "한 가지 구체예" 또는 "구체예"에 대한 본 명세서 전체를 통한 참조는 구체예와 결부되어 기재된 특별한 특징, 형태, 조성 또는 특성이 본 발명의 하나 이상의 구체예에 포함됨을 의미한다. 따라서, 본 명세서 전체에 걸친 다양한 위치에서 표현된 "한 가지 구체예에서" 또는 "구체예"의 상황은 반드시 본 발명의 동일한 구체예를 나타내지는 않는다. 추가로 특별한 특징, 형태, 조성, 또는 특성은 하나 이상의 구체예에서 어떠한 적합한 방법으로 조합될 수 있다.
Hereinafter, various embodiments described herein will be described with reference to the drawings. In the following description, for purposes of complete understanding of the present invention, various specific details are set forth, such as specific forms, compositions and processes, and the like. However, certain embodiments may be practiced without one or more of these specific details, or with other known methods and forms. In other instances, well-known processes and manufacturing techniques are not described in any detail in order not to unnecessarily obscure the present invention. Reference throughout this specification to "one embodiment" or "embodiment" means that a particular feature, form, composition, or characteristic described in connection with the embodiment is included in at least one embodiment of the invention. Accordingly, the appearances of the phrase " in one embodiment "or" an embodiment "in various places throughout this specification are not necessarily indicative of the same embodiment of the present invention. In addition, particular features, forms, compositions, or characteristics may be combined in any suitable manner in one or more embodiments.
명세서에서 특별한 정의가 없으면 본 명세서에 사용된 모든 과학적 및 기술적인 용어는 본 발명이 속하는 기술분야에서 당업자에 의하여 통상적으로 이해되는 것과 동일한 의미를 가진다.
Unless defined otherwise in the specification, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
본 발명의 일 구체예에서 “살모넬라”란, 장내세균과(Enterobacteriaceae)에 속하는 그람음성의 통성 혐기성 간균으로 S. enterica 및 S. bongori로 나뉜다. 이 2개의 종(species)중 사람과 동물에서 병원성을 발현하는 종은 S. enterica이며, 이 종은 다시 6개의 아종(subspecies; subsp.)인 enterica, salamae, arizonae, diarizoanae, houtenae 및 indica로 구분된다. 이 중에 가장 문제시 되는 아종은 S. enterica subsp. enterica이며, 이 균체들은 혈청형, 생물형 및 파지형에 따라 좀 더 세분된다. 살모넬라 균체의 표면항원은 균체항원(O-antigen), 편모항원(H-antigen) 및 표면캡슐항원(Vi-antigen)이 있으며, 카프만-화이트 분류 기준(Kauffman-White serological scheme)에 의해 균체의 혈청형이 구분되고 있다. 현재까지 알려진 혈청형은 약 2,500여 종에 달하며 그 중 200여종이 비교적 높은 발생을 보이고 있으며, 자연계에 널리 분포되어 있다.In one embodiment of the present invention, " Salmonella " is a gram-negative mixed anaerobic bacterium belonging to Enterobacteriaceae and is divided into S. enterica and S. bongori. Among these two species, S. enterica is a pathogenic organism in humans and animals. It is divided into six subspecies (subspecies): enterica, salamae, arizonae, diarizoanae, houtenae and indica do. Among the most subspecies, S. enterica subsp. enterica, which are further subdivided by serotype, biotype and phage type. The surface antigens of Salmonella strains are O-antigen, H-antigen and Vi-antigen, and they are detected by the Kauffman-White serological scheme Type. There are about 2,500 serotypes known to date, of which about 200 species are relatively high and widely distributed in nature.
살모넬라균은 숙주에 대한 적응력이 뛰어나 종 특이성이 있는 균체와 그렇게 않은 균체로 뚜렷하게 구분된다. 종 특이성이 있는 균체 혈청형 중 S. Typhi 및 S. Paratyphi는 사람에, S. Pullorum 및 S. Gallinarum은 닭에, S. Choleraesuis는 돼지에, S. Dublin은 소에, 그리고, S. Abortusovis은 양에 감염된다. 종 특이성이 낮은 혈청형에는 S. Enteritidis 및 S. Typhimurium이 대표적이며, 이 균체들은 사람을 포함한 거의 모든 동물에 감염되며 특히, 사람에게는 장염 및 패혈증을 유발시키는 인수공통병원체이다.Salmonella is highly adaptive to the host and is distinguishable from species-specific and non-species-specific strains. S. Typhi and S. Paratyphi among species-specific serotypes were found in humans, S. pullorum and S. gallinarum in chickens, S. choleraesuis in pigs, S. Dublin in cows, and S. abortusovis She is infected with sheep. S. Enteritidis and S. Typhimurium are representative serotypes with low species specificity. These strains are common pathogens that infect almost all animals including humans and cause enteritis and sepsis in humans.
살모넬라성 식중독의 오염원은 다양하지만, 그 중 계란 및 계육을 포함하는 가금 생산물은 대표적인 오염원으로 여겨지고 있으며, 이외에도 덜 익힌 육류 등 동물들의 가공품이나 부산물로 오염된 식품류에 의해서도 발생한다. 가금은 밀집 다두사육이 용이해 다른 축종보다 병원성미생물 오염기회가 높고, 질병 발생 시 제어하기가 어렵다. 오리는 닭과 달리 기본적으로 살모넬라에 저항성이 있어 주로 불현성 감염으로 나타나나, 살모넬라 감염의 매개체로 작용할 수 있다.
The source of salmonella food poisoning varies, but poultry products, including eggs and meat, are considered to be representative sources of contamination, as well as food products contaminated with processed products or by-products of animals such as less cooked meat. Poultry is easy to breed because it is more susceptible to pathogenic microorganisms than other species, and it is difficult to control when disease occurs. Ducks, unlike chickens, are basically resistant to Salmonella and appear to be predominantly infectious, but can act as mediators of Salmonella infection.
본 발명의 일 구체예에서 “저감”이란, “낮추어 줄임” 이라는 의미이다. 또한 본 명세서에서 “저감제”란, “낮추어 줄일 수 있는 제제”라는 의미로, 본 발명에 있어서의 저감제는 바람직하게 살모넬라 저감제 또는 패혈증 저감제이며, 살모넬라 감염 또는 패혈증 발병을 예방하거나, 살모넬라 감염 또는 패혈증 발병시 균의 증식을 억제하거나, 살모넬라 감염 또는 패혈증 발병시 병증을 억제할 수 있는 제제의 의미로 사용된다.
In one embodiment of the present invention, " abatement " means " lowering ". In the present specification, the term " reducing agent " means " a preparation capable of being lowered ", and the reducing agent in the present invention is preferably a salmonella reducing agent or an agent for reducing sepsis. Is used to mean an agent capable of inhibiting the growth of a microorganism at the onset of infection or sepsis, or inhibiting the disease at the onset of salmonella infection or sepsis.
본 발명의 일 구체예에서 “강황”이란, 울금이라고도 부르며 생강목에 속하는 다년생 식물로서 꽃이삭은 잎보다 먼저 나오고 넓은 달걀 모양이며 연한 녹색의 포에 싸여 있다. 4-6월에 잎겨드랑이에서 노란 꽃이 핀다. 윗부분의 포는 나비가 약간 좁고 끝은 담자홍색이며 잎겨드랑이에 꽃이 달리지 않는다. 뿌리줄기의 겉은 연한 노랑색이고, 속은 주홍빛으로 장뇌 같은 향기가 난다. 인도, 중국, 동남아시아 등지에서 많이 재배되며, 우리나라는 전남 진도, 전남 해남, 전북 부안, 경기 시흥 및 충남 청양 등지에서 재배되고 있다. 줄기와 뿌리를 식용, 약용 등으로 사용하는데, 특히 뿌리줄기를 한약재로 많이 사용한다. 맵고 쓴 맛이 나는 황색의 약재로, 통증완화와 월경불순에 효능이 있다. 인도에서는 타박상이나 염좌에 바르는 약으로 쓰며 카레 가루의 향신료로 쓰기도 한다.
In one embodiment of the present invention, " turmeric " is a perennial plant belonging to the genus Ginger, also called < RTI ID = 0.0 > Ulgum, < / RTI > Yellow flowers bloom on the leaf axils in April-June. The upper bract is slightly narrower of the butterfly, the tip is fuchsia, and the flower does not run on the leaf axil. Root stalk is light yellow on the outside and scarlet is scarlet like camphor. It is cultivated in India, China, Southeast Asia and so on. Korea is cultivated in Jindo Jeonnam, Haenam in Jeonnam, Buan in Jeonbuk, Siheung in Gyeonggi and Cheongyang in Chungnam. Stems and roots are used for edible and medicinal purposes. Root stems are often used as herbal medicines. It is a spicy, bitter, yellow medicinal product, effective for pain relief and menstrual irregularities. In India, it is used as a medicine for bruises and sprains, and also as a spice for curry powder.
본 발명의 일 구체예에서 “미강”이란, 쌀겨 또는 단순히 겨라고도 한다. 벼에서 왕겨를 뽑고 난 다음 현미를 백미로 도정하는 공정에서 분리되는 고운 속겨를 말한다. 다소의 전분이 존재하지만 지방이나 단백질이 풍부하고 인 등의 회분도 많기 때문에 주로 가축 사료에 이용된다. 도정 비율에 따라 다르지만, 대부분의 미강은 단백질 15%, 지방 17% 전후를 함유한다. 높은 함량의 지방을 포함하고 있기 때문에 헥산(hexane) 추출로 미강유를 얻기도 한다. 미강유의 원유는 적황색 또는 녹황색이지만, 탈납, 탈산, 탈취, 탈색 등의 처리를 통하여 식용유로 제조할 수 있다. 단백질은 쌀의 단백질과 비슷하고, 탄수화물은 녹말 40%, 펜토산 30% 등이다. 리파아제가 강하므로 지방은 산패되기 쉽다. 또 비타민 B1은 2mg% 이상 함유된다.In one embodiment of the present invention, " rice bran " is also called rice bran or simply bran. It is a fine puke that is separated from the process of removing rice hulls from rice and then turning brown rice into white rice. There are some starches, but they are mainly used for livestock feed because they are rich in fat and protein, and there are many ashes such as phosphorus. Most of the rice bran contains about 15% protein and 17% fat, depending on the ratio. Because it contains a high fat content, hexane extraction can be used to obtain rice bran oil. Although crude oil of rice bran oil is reddish yellow or greenish yellow, it can be produced from edible oil through treatment such as dehydration, deoxidation, deodorization, discoloration. Protein is similar to protein in rice, carbohydrate is 40% starch, 30% pentanoic acid. The fat is prone to sowing because of the strong lipase. Vitamin B1 is more than 2mg%.
본 명세서에 있어서, 미강은 현미를 백미로 도정하는 공정에서 분리되는 하얀 빛깔의 쌀겨를 의미하여, 표면이 검은 빛깔을 나타내는 흑미강과 구분된다.
In the present specification, rice bran means rice bran of white color separated in the process of converting brown rice into white rice, and the surface is distinguished from black rice which shows black color.
본 발명의 일 구체예에서 “산초”란, 쌍떡잎식물 쥐손이풀목 운향과의 낙엽관목으로 산지에서 자란다. 높이는 3m이고 잔가지는 가시가 있으며 붉은빛이 도는 갈색이다. 잎은 어긋나고 13-21개의 작은잎으로 구성된 깃꼴겹잎이다. 작은잎은 길이 1-5cm의 넓은 바소꼴이며 양끝이 좁고 가장자리에 물결 모양의 톱니와 더불어 투명한 유점이 있다. 산초는 잎을 따서 생으로 사용하거나, 열매껍질을 벗겨 말린 후 달이거나 농축해서 사용한다. 산초 열매는 소화불량, 식체, 위하수, 위확장, 구토, 이질, 설사, 신경쇠약, 기침, 회충 구제 등에 효능이 있다. 요리에는 후추와 비슷한 방법으로 사용된다.
In one embodiment of the present invention, " Sancho " is a deciduous shrub with a dicotyledonous mushroom, and grows in a mountainous area. It is 3m high and twigs are spiny and reddish brown. Leaves are alternate phyllotaxis of 13-21 small leaves. The small leaf is a wide basaloid with a length of 1-5cm, narrow both ends, with wavy sawtooth on the edge, and a transparent oil spot. Pickled leaves are used as raw leaves, dried or peeled, and then dried or concentrated. Sancho fruit is effective for indigestion, diarrhea, gastric warts, gastric enlargement, vomiting, diarrhea, diarrhea, nervous breakdown, cough, and redness. Cooking is used in a similar way to pepper.
본 발명의 일 구체예에서 “담자균류”란, 진균류의 한 아문으로 유성생식 한 결과로 담자기라는 세포가 되어 홀씨를 만드는 균류이다. 스스로 양분을 만들지 못하여 다른 생물체에 붙어 기생하며, 버섯으로 알려진 것이 많은데 상황버섯, 차가버섯, 표고버섯, 잎새버섯, 목이버섯, 눈꽃송이버섯 및 치마버섯 따위가 대표적이다. 성숙한 버섯의 갓주름 면에는 담자기가 빽빽하게 생긴다. 담자균류 버섯의 대부분은 고등식물의 유체에서 부생적으로 생활하며, 특히 셀룰로오스와 리그닌을 분해하여 물질의 생물학적 순환에 없어서는 안 될 중요한 역할을 하고 있으며, 또 수목의 뿌리에 균근을 형성하여 공동생활을 하는 것도 많다. 그밖에 식용균으로 중요시되거나 독버섯으로 주의를 해야 할 종류도 있다.
In one embodiment of the present invention, " basidiomycetes " is a fungus that forms a cell as a result of sexual reproduction of a fungus as a result of sexual reproduction. There are many things known as mushrooms which are parasitic to other organisms because they can not make their own nutrients. Typical examples are mushroom, mushroom, shiitake, mushroom, mushroom, and mushroom. On the freshly corrugated surface of mature mushrooms, a thick layer of volcanic ash is formed. Most of the mushrooms live as secondary organisms in the fluid of higher plants. In particular, they decompose cellulose and lignin and play an important role in the biological circulation of the material. In addition, they form mycorrhiza in the roots of trees, There are many things to do. There are other kinds that are important as edible bacillus or should be cautious with poisoning.
본 발명의 일 구체예에서 “균사”란, 사상균(곰팡이)의 영양세포이다. 영양적 기능이나 생리적 기능 같은 기능 분화도 볼 수 있다. 형태적으로는 격벽의 유무가 분류에 사용되고 있으며, 포자의 발아관에서 발육하여 선단생장에 의해서 신장한다. 분지한 균사의 집단을 균사체(mycelium)라고 한다. 생리적 기능에 따라 배지나 기주의 표면 또는 내부에 들어가면서 신장하는 기저균사와 공기 중에서 신장하는 기생균사로 구별된다. 기생균사는 영양을 기저균사로부터의 이송에 의존하고 있으며, 그 일부는 포자 형성기관 등으로 분화한다. 유기물이 있고 적당한 습도와 온도 등 환경조건이 양호하면 여러 가지 균사조직을 형성한다. 자낭균류의 균사의 격막은 단순하지만 담자균류에서는 꺾쇠연결을 형성하는 것을 많이 볼 수 있다. 곰팡이류에서는 여러 갈래로 분지하여 실처럼 엉겨 있고, 버섯류에서는 집합하여 자실체를 만든다. 엽록소가 없어 광합성을 하지 못하고 기주생물에 붙어서 영양분을 흡수한다. 버섯인 담자균류의 경우, 포자에서 발아한 균사는 핵을 1개 가지고, 이것이 다른 균사와 융합하면 1개의 세포에 핵을 2개 가지게 된다. 핵을 1개 가진 균사를 1차 균사, 핵을 2개 가진 균사를 2차 균사라고 한다. 균사의 세포벽은 일반적으로 키틴(chitin) 또는 헤미셀룰로오스(hemicellulose)가 주성분이며 이 둘을 합한 경우도 있다.In one embodiment of the present invention, " mycelium " is a nutritive cell of a filamentous fungus (mold). Functional differentiation such as nutritional function or physiological function can also be seen. Morphologically, the presence or absence of the bulkhead is used for classification, and it develops in germination tube of spore and elongates by tip growth. The group of branched mycelium is called mycelium. Depending on the physiological function, it is distinguished into basal hypha that grows on the surface or inside of the pouch or host, and parasitic bacteria which grows in the air. Parasitoids depend on the transfer of nutrients from basal hyphae, some of which differentiate into spore forming organs and the like. If there are organic matter and suitable environmental conditions such as appropriate humidity and temperature, various mycelial tissues are formed. The mycelial membrane of mycelium of fungus is simple, but it can be seen that it forms a braid connection. In the fungi, it branches into several branches and is entangled like a thread. It does not have chlorophyll and does not have photosynthesis. It attaches to the host organism and absorbs nutrients. In the case of the mushroom-bearing bacterium, the mycelium germinated in the spore has one nucleus, which when fused with another mycelium has two nuclei in one cell. Mycelium with one nucleus is called mycelium, and mycelium with two nuclei is called secondary mycelium. The cell wall of the mycelium is generally composed of chitin or hemicellulose, which is sometimes combined.
본 명세서에 있어서 담자균류균사는 구체적으로 약용 및 식용 가능한 담자균류인 상황버섯, 차가버섯, 표고버섯, 잎새버섯, 목이버섯, 눈꽃송이버섯, 치마버섯 등의 균사에서 어느 하나를 선택하여 사용할 수 있으며, 이들 담자균류 균사는 한국미생물보존센터(KCCM), 한국생명공학연구원 생명자원센터(KCTC), 한국세포주은행(KCLB), 한국농업미생물자원센터(KACC), 미국표준균주배양수록보존소(ATCC) 등의 기관에서 균주를 분양 받아 사용할 수 있다.
In the present specification, the mycelia of mycelia can be selected from mycelium of mushroom, chaga mushroom, shiitake mushroom, mushroom mushroom, mushroom mushroom, The mycelia of these mycelia were collected from the KCCM, KCTC, KCLB, KACC, ATCC, And the like.
본 발명의 일 구체예에서 “생물전환”이란, 생물의 생리적 기능을 이용해 첨가된 물질을 화학적으로 변형된 형태로 전환시키는 것을 말한다.
In one embodiment of the present invention, " bioconversion " refers to the conversion of a substance added using a physiological function of an organism into a chemically modified form.
본 발명의 일 구체예에서 “발효”란, 미생물이 자신이 가지고 있는 효소를 이용해 유기물을 분해시키는 과정을 말한다. 발효라고 하는 것은 효소작용에 의해 유기물이 간단한 화합물로 변화해 자유 에너지를 내놓는 현상이지만, 일반적으로는 미생물이 유기물을 분해해서 대사물을 축적하는 현상을 말한다. 즉, 효모가 당을 무산소적으로 분해해서 에틸알코올과 이산화탄소로 하는 알코올발효, 락트산균이 당을 무산소적으로 분해해서 락트산으로 하는 락트산발효 등이 전형적인 발효이지만, 현재는 아세트산균이 공기 중의 산소를 이용해서 알코올을 아세트산으로 산화시키는 현상, 곰팡이가 공기 중의 산소를 이용해서 글루코오스를 글루콘산으로 산화시키는 현상 등도 각각 아세트산발효, 글루콘산발효 등으로 부르고 있다. 또 이러한 것들을 혐기적 발효, 호기적 발효(산화 발효)로 나누어 논하기도 하고, 상기 언급한 것 외의 혐기적 발효로는 글리세롤발효, 아세톤부탄올발효, 2, 3-부틸렌글리콜발효, 부티르산발효, 프로피온산발효, 메탄발효 등이, 또 호기적 발효로는 시트르산발효, 이타콘산발효, 효모산발효, 2-케토글루타르산발효, 옥살산발효, 푸마르산발효, 소르보오스발효 등이 있다. 이외에 미생물에 의한 아미노산, 비타민, 항생 물질의 생산도 예를 들면 글루탐산발효, 리보플라빈발효, 페니실린발효 등으로 불리는데, 그다지 좋은 호칭은 아니다. 발효 작용명은 생산물로 부르는 것이 원칙이나, 때로는 셀룰로오스발효, 펙틴발효 등으로 기질의 이름을 붙이기도 한다.
In one embodiment of the present invention, " fermentation " refers to a process in which a microorganism decomposes an organic substance using an enzyme contained therein. Fermentation is a phenomenon in which organic matter changes into a simple compound by enzymatic action and releases free energy. In general, microorganisms decompose organic matter and accumulate metabolites. That is, fermentation of alcohol by anaerobic digestion of sugars by anaerobic digestion of yeast, fermentation of lactic acid by lactic acid bacteria decomposing sugar nonoxiously into lactic acid is a typical fermentation, but now, A phenomenon in which alcohol is oxidized to acetic acid by use of oxygen, and a phenomenon in which mold oxidizes glucose to gluconic acid by using oxygen in the air are also referred to as acetic acid fermentation and gluconic acid fermentation. These anaerobic fermentation can be divided into anaerobic fermentation and aerobic fermentation (oxidation fermentation), and other anaerobic fermentation furnaces include glycerol fermentation, acetone butanol fermentation, 2, 3-butylene glycol fermentation, butyric acid fermentation, Fermentation, and methane fermentation. Examples of aerobic fermentation include citric acid fermentation, itaconic acid fermentation, yeast acid fermentation, 2-keto glutaric acid fermentation, oxalic acid fermentation, fumaric acid fermentation and sorbose fermentation. In addition, production of amino acids, vitamins and antibiotics by microorganisms is also called glutamic acid fermentation, riboflavin fermentation, penicillin fermentation and the like, which is not a good title. The name of the fermentation is usually called the product, but sometimes the name of the substrate is given by cellulosic fermentation, pectin fermentation and so on.
본 발명의 일 구체예에서 “가금류”란, 고기, 알, 털, 깃털 등 인간에게 유용한 축산물을 얻기 위해서 사람에 의해 사육되는 조류를 의미하며, 이에 한정하지는 않지만, 예를 들어 닭(육계), 오리, 칠면조, 거위, 화조, 집비둘기 및 메추리 등을 말한다. 특히, 육계는 가금류에 속하는 다른 조류와 알에서 부화되는 특징, 날개와 부리를 가지고 있는 특징, 몸이 깃털로 덮이는 특징 및 폐호흡을 하며 척추를 가지는 특징의 공통점이 있다는 점에서, 육계의 생육과 관련된 효과는 기타 다른 가금류에도 그 효과가 있다고 할 것이다. 또한 본 명세서에 있어서, 가금류용 사료란 가금류의 양적, 질적 크기를 변화시키기 위해 공급하는 식용 물질을 말한다. 여기에 함유되는 발효 조성물은, 농축액 또는 분말 등의 형태 변화를 포함한다.
In one embodiment of the present invention, the term " poultry " means a bird raised by a person in order to obtain livestock products useful for human beings such as meat, eggs, hairs and feathers. Ducks, turkeys, geese, ancestors, pigeons and quails. In particular, broiler chickens have the characteristics of hatching in other birds and eggs of poultry, the characteristics of having wings and beaks, the features of the body being covered by feathers, Will be effective for other poultry. In this specification, feed for poultry refers to edible material supplied to change the quantitative and qualitative size of poultry. The fermentation composition contained therein includes morphological changes such as a concentrate or powder.
본 발명의 일 구체예에서 “패혈증”이란, 미생물에 감염되어 전신에 심각한 염증반응이 나타나는 상태를 말한다. 체온이 38도 이상으로 올라가는 발열증상 혹은 36도 이하로 내려가는 저체온증, 호흡수가 분당 24회 이상으로 증가(빈호흡), 분당 90회 이상의 심박수(빈맥), 혈액검사상 백혈구 수의 증가 혹은 현저한 감소 중 두 가지 이상의 증상을 보이는 경우, 이를 전신성 염증반응 증후군(systemic inflammatory response syndrome; SIRS)이라고 부른다. 이러한 전신성 염증반응 증후군이 미생물의 감염에 의한 것일 때 패혈증이라고 한다. 원인이 되는 감염 부위는 신체의 모든 장기가 가능하다. 폐렴, 신우신염, 뇌막염, 봉와직염, 감염성 심내막염, 복막염, 욕창, 담낭염, 담도염 등이 패혈증의 원인이 될 수 있다. 이러한 감염증이 발생한 경우, 원인미생물이 혈액 내로 침범하여 패혈증을 일으킬 수 있다. 그러나 미생물이 혈액 내로 침투하지 않더라도 신체 일부의 염증 반응 및 염증물질의 생성에 의해서 전신적인 패혈증이 발생할 수도 있다. 패혈증의 초기 증상으로는 호흡수가 빨라지고, 지남력(시간, 장소, 사람에 대한 인지력)의 상실이나 정신착란 등의 신경학적 장애가 나타날 수 있다. 혈압의 저하 및 신체 말단에 공급되는 혈액량의 저하로 인하여 피부가 시퍼렇게 보이기도 한다. 균혈증(세균이 혈액 내에 돌아다니는 증상)이 있으면 세균이 혈류를 따라 돌아다니다가 신체의 특정 부위에 자리를 잡아 그 부위에 병적인 변화를 일으킬 수 있다. 원인균에 특이적인 피부의 변화가 나타나므로, 패혈증의 원인을 진단하는 데 도움이 될 수 있다. 소화기 계통의 증상으로는 구역, 구토, 설사 및 장 마비 증세가 나타나고 심한 스트레스 상황에서는 소화기의 출혈증상도 나타날 수 있다.
In one embodiment of the present invention, " sepsis " refers to a condition in which a microorganism is infected and a serious inflammatory reaction occurs in the whole body. The number of breaths increased more than 24 times per minute (ventilation), heart rate more than 90 times per minute (tachycardia), increase in leukocyte count in blood test, or significant decrease in respiratory rate If you have symptoms that are more than a branch, it is called systemic inflammatory response syndrome (SIRS). This systemic inflammatory response syndrome is called sepsis when it is caused by microbial infection. The source of the infection can be any organs of the body. Pneumonia, pyelonephritis, meningitis, cellulitis, infective endocarditis, peritonitis, pressure ulcer, cholecystitis, and cholangitis may cause sepsis. If such infection occurs, the causative microorganism may invade into the blood and cause sepsis. However, even if microorganisms do not penetrate into the blood, systemic sepsis may occur due to the inflammation reaction of some body parts and the production of inflammatory substances. Early symptoms of sepsis may include respiratory arrest, neurological disturbances such as loss of locomotion (time, place, cognitive ability) or delirium. The skin may appear visually opaque due to lowering of blood pressure and lowering of the amount of blood supplied to the distal end of the body. Bacteremia (a symptom of germs circulating in the blood) can cause germs to move around the bloodstream, to position themselves in certain parts of the body and cause pathological changes in those areas. As the skin changes specific to the causative bacteria, it may help to diagnose the cause of sepsis. Symptoms of the digestive system include nausea, vomiting, diarrhea and intestinal paralysis, and severe gastrointestinal bleeding may also occur.
본 발명의 일 구체예에서 “약학조성물”이란, 특정한 목적을 위해 투여되는 조성물을 의미한다. 본 발명의 목적상, 본 발명의 약학조성물은 살모넬라 감염 또는 패혈증 발병군에서 생존율을 증진시키거나, 염증활성 물질의 억제를 위해 사용되는 것이며, 이를 위한 조성물 및 약학적으로 허용 가능한 담체, 부형제 또는 희석제를 포함할 수 있다. 본 발명에 따른 약학 조성물은, 조성물 총 중량에 대하여 본 발명의 약학조성물에 상응하는 유효성분을 0.1 내지 50 중량%로 포함한다. 본 발명의 약학조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다.
&Quot; Pharmaceutical composition " in one embodiment of the present invention means a composition to be administered for a specific purpose. For the purpose of the present invention, the pharmaceutical composition of the present invention is used for improving the survival rate or inhibiting the inflammatory active substance in a group infected with Salmonella or sepsis, and a composition therefor and a pharmaceutically acceptable carrier, excipient or diluent . ≪ / RTI > The pharmaceutical composition according to the present invention comprises 0.1 to 50% by weight of the active ingredient corresponding to the pharmaceutical composition of the present invention, based on the total weight of the composition. 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 phosphate, calcium silicate , Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
본 발명의 일 구체예에서 “식품조성물”이란, 살모넬라 감염 또는 패혈증 질환의 개선을 위한 식품조성물로 다양하게 이용되는 것으로서, 본 발명의 조성물을 유효성분으로 포함하는 식품조성물은 각종 식품류, 예를 들어, 음료, 껌, 차, 비타민 복합제, 분말, 과립, 정제, 캡슐, 과자, 떡, 빵 등의 형태로 제조될 수 있다. 본 발명의 식품조성물은 독성 및 부작용이 거의 없는 천연식품 및 이의 발효물로 구성된 것이므로 예방 목적으로 장기간 복용 시에도 안심하고 사용할 수 있다. 본 발명의 조성물이 식품조성물에 포함될 때 그 양은 전체 중량의 0.1 내지 50%의 비율로 첨가할 수 있다. 여기서, 상기 식품조성물이 음료 형태로 제조되는 경우 지시된 비율로 상기 식품조성물을 함유하는 것 외에 특별한 제한점은 없으며 통상의 음료와 같이 여러가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 즉, 천연 탄수화물로서 포도당 등의 모노사카라이드, 과당 등의 디사카라이드, 슈크로스 등의 및 폴리사카라이드, 덱스트린, 시클로덱스트린 등과 같은 통상적인 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜 등을 포함할 수 있다. 상기 향미제로서는 천연 향미제(타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진등) 및 합성 향미제(사카린, 아스파르탐 등) 등을 들 수 있다. 그 외 본 발명의 식품조성물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 그렇게 중요하진 않지만 본 발명의 조성물 100 중량부 당 0.1 내지 약 50 중량부의 범위에서 선택되는 것이 일반적이다.
In one embodiment of the present invention, the term " food composition " is used variously as a food composition for improving salmonella infection or sepsis disease. The food composition comprising the composition of the present invention as an active ingredient may be used in various foods, , Beverages, gums, tea, vitamin complexes, powders, granules, tablets, capsules, confectionery, rice cakes and breads. Since the food composition of the present invention is composed of a natural food and its fermented product which have little toxicity and side effects, it can be safely used for prolonged use even for prophylactic purposes. When the composition of the present invention is contained in the food composition, the amount thereof may be added in a proportion of 0.1 to 50% of the total weight. Here, when the food composition is prepared in a beverage form, there are no particular limitations other than those containing the food composition at the indicated ratios and may contain various flavors or natural carbohydrates such as ordinary beverages as an additional ingredient. That is, natural carbohydrates include monosaccharides such as glucose, disaccharides such as fructose, sucrose and the like, and sugar sugars such as polysaccharide, dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol can do. Examples of the flavors include natural flavors (such as tau martin, stevia extract (for example, rebaudioside A and glycyrrhizin), and synthetic flavors (for example, saccharin and aspartame). The food composition of the present invention can be used as a food composition containing various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants, pectic acid and its salts, alginic acid and its salts, Stabilizers, antiseptics, glycerin, alcohols, carbonating agents used in carbonated beverages, etc. These components may be used independently or in combination. The ratio of these additives is not so important, Is generally selected in the range of 0.1 to about 50 parts by weight per part.
본 발명의 일 구체예에서 “사료조성물”이란, 시판되고 있는 일반사료를 모두 포함할 수 있으며, 이에 한정되지는 않으나, 미강, 옥수수, 대두박, 콩, 수수, 쌀, 보리, 밀, 귀리, 호밀, 좁쌀, 메밀, 티리티게일, 고구마, 타피오카, 밀기울, 맥강, 대두피, 옥수수겨, 맥아근, 전분박, 커피박, 잠분, 잠사, 해조분, 면실박, 채종박, 캐놀라밀, 임자박, 호마박, 아마박, 해바라기씨박, 낙화생박, 야자박, 옥수수 글루텐, 주정박, 옥수수 배아박, 고추씨박, 루핀종실, 어분, 우모분 및 육분일 수 있으며, 이러한 사료에 상기의 담자균류 발효 조성물을 함유하는 것을 포함한다. 본 발명의 사료에는 일반적인 사료 첨가제, 예를 들면 소금, 골분, 인산칼슘제, 무기물 혼합제, 비타민제, 아미노산제, 항생물질, 호르몬제 등의 특수목적을 위한 사료 첨가제가 포함될 수 있다.
In one embodiment of the present invention, the term " feed composition " includes all commercially available general diets, including but not limited to rice bran, corn, soybean meal, soybean meal, rice, barley, wheat, oats, rye Corn bran, starch powder, coffee starch, cloves, clam, seaweed, cottonseed, seeds, canola wheat, chestnut, wheat, barley, Corn gluten meal, corn germ meal, red pepper seed meal, lupine seed meal, fish meal, oyster meal and meat meal, and these feeds may be subjected to fermentation ≪ / RTI > compositions. Feeds of the present invention may include feed additives for general purpose such as general feed additives such as salt, bone meal, calcium phosphate, mineral compound, vitamin, amino acid, antibiotic, hormone and the like.
본 발명의 일 구체예에서, (a) 미강을 셀룰라아제 및 헤미셀룰라아제 계열의 효소와 아밀라아제 계열의 효소로 가수분해하는 단계; (b) 상기 (a)단계에서의 가수분해물에 담자균류균사를 접종하고, 배양하여 발효물을 제조하는 단계; 및 (c) 상기 (b)단계에서의 배양공정에 의해 생산된 발효물에 섬유소분해효소를 처리하는 단계를 포함하는 미강생물전환산물의 제조방법을 제공하고, 상기 담자균류균사는 상황버섯, 차가버섯, 표고버섯, 잎새버섯, 목이버섯, 눈꽃송이버섯, 치마버섯으로 이루어진 그룹으로부터 선택된 어느 하나 이상인 미강생물전환산물의 제조방법을 제공하며, 상기 (c)단계에서의 섬유소분해효소는 세포벽 구성성분을 분해할 수 있는 효소인 것인 미강생물전환산물의 제조방법을 제공하며, 상기 섬유소분해효소는 셀룰라아제, 헤미셀룰라아제, 펙티나아제 및 글루카나제로 이루어진 그룹으로부터 선택된 어느 하나 이상인 미강생물전환산물의 제조방법을 제공한다.
In one embodiment of the present invention, there is provided a method for producing a microorganism, comprising the steps of: (a) hydrolyzing a rice germ into a cellulase and a hemicellulase family of enzymes and an amylase family of enzymes; (b) inoculating the hydrolyzate of step (a) with mycelia of mycelia and culturing to produce a fermented product; And (c) treating the fermented product produced by the culturing step in the step (b) with a fibrinolytic enzyme, wherein the mycelium of the bacterium belongs to the mushroom, Wherein the cellulolytic enzyme in step (c) comprises at least one member selected from the group consisting of mushrooms, shiitake mushrooms, mushrooms, mushrooms, snow mushrooms, Wherein the cellulolytic enzyme is at least one selected from the group consisting of cellulase, hemicellulase, pectinase, and glucanase. ≪ / RTI >
본 발명의 다른 구체예에서, 미강을 셀룰라아제 및 헤미셀룰라아제 계열의 효소와 아밀라아제 계열의 효소로 가수분해하고 담자균류균사를 접종하여 배양한 발효물에 섬유소분해효소를 처리한 미강생물전환산물을 유효성분으로 포함하는 약학조성물을 제공하고, 상기 약학조성물은 살모넬라 감염의 예방 또는 치료 목적인 약학조성물을 제공하며, 상기 약학조성물은 살모넬라에 대한 대식세포의 탐식능 및 탐식 후 살해능을 향상시키는 약학조성물을 제공하며, 상기 약학조성물은 살모넬라 감염시 Th1 도움 T세포(Th1 helper T cell) 면역반응 유도를 향상시키는 약학조성물을 제공하며, 상기 약학조성물은 패혈증의 예방 또는 치료 목적인 약학조성물을 제공한다.
In another embodiment of the present invention, the rice bran bioconversion product obtained by hydrolyzing rice bran with enzymes of the cellulase and hemicellulase family enzymes and the amylase family enzymes, inoculating the mycelia with the mycelium and treating the fermented product with the fibrinolytic enzyme, Wherein said pharmaceutical composition provides a pharmaceutical composition for the prevention or treatment of Salmonella infection, said pharmaceutical composition providing a pharmaceutical composition for enhancing the phagocytosis and phagocytosis of macrophages against Salmonella, , The pharmaceutical composition provides a pharmaceutical composition for improving the induction of Th1 helper T cell immune response upon Salmonella infection, wherein the pharmaceutical composition provides a pharmaceutical composition for the prevention or treatment of sepsis.
본 발명의 또 다른 구체예에서, 미강을 셀룰라아제 및 헤미셀룰라아제 계열의 효소와 아밀라아제 계열의 효소로 가수분해하고 담자균류균사를 접종하여 배양한 발효물에 섬유소분해효소를 처리한 미강생물전환산물을 유효성분으로 포함하는 식품조성물을 제공한다.
In another embodiment of the present invention, a rice bran bioconversion product obtained by hydrolyzing rice bran with an enzyme of a cellulase and hemicellulase family enzymes and an amylase family of enzymes, inoculating a mycelium of mycelia with a mycelium, ≪ / RTI >
본 발명의 또 다른 구체예에서, 미강을 셀룰라아제 및 헤미셀룰라아제 계열의 효소와 아밀라아제 계열의 효소로 가수분해하고 담자균류균사를 접종하여 배양한 발효물에 섬유소분해효소를 처리한 미강생물전환산물을 유효성분으로 포함하는 사료조성물을 제공하고, 상기 사료조성물은 소, 돼지, 가금류 등의 사육용인 사료조성물을 제공하며, 상기 가금류는 닭, 오리, 칠면조, 거위, 화조, 집비둘기 및 메추리 등으로 구성된 군으로부터 선택된 어느 하나 이상인 사료조성물을 제공한다.
In another embodiment of the present invention, a rice bran bioconversion product obtained by hydrolyzing rice bran with an enzyme of a cellulase and hemicellulase family enzymes and an amylase family of enzymes, inoculating a mycelium of mycelia with a mycelium, Wherein the feed composition provides feed compositions for cattle such as cows, pigs, poultry and the like, wherein the poultry is selected from the group consisting of chicken, duck, turkey, goose, At least one selected feed composition is provided.
이하 상기 본 발명을 단계별로 상세히 설명한다.
Hereinafter, the present invention will be described in detail.
미생물(담자균류균사) 발효 및 효소처리 생물전환공정을 통해 생물전환된 미강생물전환산물은 소, 돼지, 및 가금류를 포함한 가축의 살모넬라 감염 및 패혈증 발병 저감에 큰 효과가 있으므로, 안전한 축산물 생산에 크게 기여할 것으로 기대된다.
Fermentation and Enzyme Treatment of Microorganisms (Mycelial Microflora) Fermentation and Biological Conversion of Biological Converted Biogas Conversion Products are effective in reducing the incidence of salmonella infection and sepsis in livestock including cattle, pigs, and poultry, It is expected to contribute.
도 1은 본 발명의 일 실시예에 따른, 강황, 미강, 또는 산초 원재료의 생물전환공정 및 생물전환산물 분말 수득 과정을 나타낸 모식도이다.
도 2는 본 발명의 일 실시예에 따른, 강황, 미강, 또는 산초 생물전환산물이 처리된 대식세포에서의 살모넬라 탐식능 및 살해능 향상 결과를 나타낸 그래프이다.
도 3은 본 발명의 일 실시예에 따른, 강황 원산지별 원물 또는 생물전환산물의 대식세포 살모넬라 탐식능 향상 결과를 나타낸 그래프이다.
도 4는 본 발명의 일 실시예에 따른, 강황 원산지별 원물 또는 생물전환산물의 대식세포 살모넬라 살해능 향상 결과를 나타낸 그래프이다.
도 5는 본 발명의 일 실시예에 따른, 치사량 살모넬라 감염 마우스 모델에서의 생물전환산물 복강투여시 마우스 생존율 향상 효능 평가 결과를 나타낸 그래프이다.
도 6은 본 발명의 일 실시예에 따른, 치사량 살모넬라 감염 마우스 모델에서의 생물전환산물 경구투여시 마우스 생존율 향상 효능 평가 결과를 나타낸 그래프이다.
도 7은 본 발명의 일 실시예에 따른, 치사량 살모넬라 감염 마우스 모델에서의 생물전환산물 복강투여 및 경구투여시 마우스 생존율 향상 효능 평가 결과를 나타낸 그래프이다.
도 8은 본 발명의 일 실시예에 따른, 아치사량 살모넬라 감염 마우스 모델에서의 생물전환산물 투여시 마우스 복강 내 살모넬라 집락수 계측 결과를 나타낸 그래프이다.
도 9는 본 발명의 일 실시예에 따른, 아치사량 살모넬라 감염 마우스 모델에서의 생물전환산물 투여시 마우스 비장 내 림프구수 계측 결과를 나타낸 그래프이다.
도 10은 본 발명의 일 실시예에 따른, 아치사량 살모넬라 감염 마우스 모델에서의 생물전환산물 투여시 IFN-γ, IL-1β, IL-2 및 IL-5 사이토카인의 발현량 변화를 나타낸 그래프이다.
도 11은 본 발명의 일 실시예에 따른, 아치사량 살모넬라 감염 마우스 모델에서의 강황 생물전환산물 투여시 IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10 및 IL-12 사이토카인의 발현량 변화를 나타낸 그래프이다.
도 12는 본 발명의 일 실시예에 따른, 아치사량 살모넬라 감염 마우스 모델에서의 미강 생물전환산물 투여시 IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10 및 IL-12 사이토카인의 발현량 변화를 나타낸 그래프이다.
도 13은 본 발명의 일 실시예에 따른, 아치사량 살모넬라 감염 마우스 모델에서의 강황 또는 미강의 생물전환산물 투여시 간 조직의 손상 억제 효과를 나타낸 도이다.
도 14는 본 발명의 일 실시예에 따른, LPS 유도 패혈증 모델에서의 생물전환산물 투여 농도에 따른 마우스 생존율 향상 효능 평가 결과를 나타낸 그래프이다.
도 15는 본 발명의 일 실시예에 따른, LPS 유도 패혈증 모델에서의 생물전환산물 투여 경로에 따른 마우스 생존율 향상 효능 평가 결과를 나타낸 그래프이다.
도 16은 본 발명의 일 실시예에 따른, LPS 유도 패혈증 모델에서의 동일 투여 농도 및 투여방법 기준 생물전환산물별 마우스 생존율 차이를 나타낸 그래프이다.
도 17은 본 발명의 일 실시예에 따른, 미강과 흑미강의 생물전환산물의 대식세포에 대한 면역자극효과를 NO(Nitric Oxide) 분석 결과로 나타낸 그래프이다.FIG. 1 is a schematic view showing a biological conversion process of a raw material of turmeric, rice bran, or anthraquinone and a process of obtaining a biotransformation product powder according to an embodiment of the present invention.
FIG. 2 is a graph showing the results of salmonella sputum smearing and killing enhancement in macrophages treated with turmeric, rice bran, or sancho bioconversion products according to an embodiment of the present invention.
FIG. 3 is a graph showing the results of improving the macrophage Salmonella sputum production ability of raw or biotransformed products according to one embodiment of the present invention.
FIG. 4 is a graph showing the results of improving the viability of macrophages Salmonella in native or bioconverted products according to one embodiment of the present invention.
FIG. 5 is a graph showing the results of evaluating the effect of improving the survival rate of mice in the case of intraperitoneal administration of biotransformation products in a mouse model of lethal Salmonella infection, according to an embodiment of the present invention.
FIG. 6 is a graph showing the results of evaluating the effect of improving the survival rate of mice when the biotransformation product is orally administered in a mouse model of lethal Salmonella infection according to an embodiment of the present invention.
FIG. 7 is a graph showing the results of evaluating the effect of improving the survival rate of mice in the case of oral administration of the biotransformation product in the lethal dose Salmonella-infected mouse model according to an embodiment of the present invention.
FIG. 8 is a graph showing the results of measurement of Salmonella colonies in mouse abdominal cavity during administration of a biotransfer product in a mouse model of asthma salmonella infection according to an embodiment of the present invention.
FIG. 9 is a graph showing the results of measurement of lymphocyte count in mouse spleen upon administration of a biotransfer product in a mouse model of asthma Salmonella infection according to an embodiment of the present invention. FIG.
Figure 10 is a graph showing changes in expression levels of IFN-y, IL-l [beta], IL-2 and IL-5 cytokines upon administration of biotransformation products in a mouse model of asthma Salmonella infection in accordance with an embodiment of the present invention .
IL-1, IL-2, IL-4, IL-5, IL-6, IL-6, and IL-6, respectively, upon administration of a turmeric biotransformation product in a mouse model of infection with Salmonella typhimurium according to an embodiment of the present invention. IL-10 < / RTI > and < RTI ID = 0.0 > IL-12 < / RTI > cytokines.
IL-4, IL-5, IL-6, IL-6, and IL-10 in the mouse model of infectious salmonellosis infection according to an embodiment of the present invention. IL-10 < / RTI > and < RTI ID = 0.0 > IL-12 < / RTI > cytokines.
FIG. 13 is a graph showing an effect of inhibiting damage to liver tissues upon administration of a biotransfer product of turmeric or rice bran in a mouse model of saliva Salmonella infection in accordance with an embodiment of the present invention.
FIG. 14 is a graph showing the results of evaluating the effect of improving the survival rate of mice according to the concentration of the bioconversion product in the LPS-induced sepsis model according to an embodiment of the present invention.
15 is a graph showing the results of evaluating the effect of improving the survival rate of mice according to the route of biotransformation products in the LPS-induced sepsis model according to an embodiment of the present invention.
FIG. 16 is a graph showing the mouse survival rate difference according to the same administration dose and administration method based on the biotransformation product in the LPS-induced sepsis model according to an embodiment of the present invention.
FIG. 17 is a graph showing NO (nitric oxide) analysis results on the immunostimulatory effect of macrophages on the macrophages of biotransformation products of rice bran and black rice, according to an embodiment of the present invention.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을보다 구체적으로 설명하기 위한 것으로서, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.
Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .
실시예 1. 발효 및 효소처리의 생물전환공정Example 1 Bioconversion Process of Fermentation and Enzyme Treatment
실시예 1-1. 원재료의 전처리Example 1-1. Pretreatment of raw materials
구입한 강황(대한민국 진도산, 인도산), 미강, 산초의 원재료 전처리 작업을 위하여 원재료 상태의 곰팡이 오염도를 측정한 후, 천연물 원재료의 이물 및 오염물 제거를 위한 세척작업을 하였다. 세척작업은 총 3단계를 거쳐 수행하였으며 ①이물 및 곰팡이 포자를 제거하기 위한 공기세척 단계, ②잔류 농약 등의 제거를 위한 물세척 단계, ③미생물 등의 오염을 제거하기 위한 알코올(Et-OH)세척 단계를 통해 원재료의 세척작업을 수행하였다. 세척 및 절단작업을 마친 원재료 각각은 대량건조가 가능한 열풍건조기에서 건조작업을 수행한 후 조분쇄 및 미분쇄 공정을 거쳐 각각의 천연물 원재료의 분말화 작업을 수행하였다.
In order to pre-treat the raw materials of purchased ganghwang (Jindo, Indo), rice gruel, and sancho, the mold contamination degree of the raw material was measured and then washing work was performed to remove foreign matters and contaminants of the natural raw materials. (1) air washing step to remove foreign matters and mold spores, (2) water washing step to remove residual pesticides, (3) alcohol (Et-OH) to remove contaminants such as microorganisms, The washing of the raw materials was carried out through the washing step. Each of the raw materials after washing and cutting was dried in a hot air drier capable of mass drying, and subjected to pulverization of raw materials of raw materials through crude pulverization and pulverization.
실시예 1-2. 건조재료의 생물전환산물 수득Examples 1-2. Obtain bioconversion products of dry materials
이물 및 곰팡이 오염을 제거한 강황, 미강, 또는 산초 각각은 액상 배양배지화를 위해 효소처리 및 열처리 살균공정을 수행하였다. 효소는 나무껍질, 근피, 뿌리, 잎 등을 분해할 수 있는 셀룰라아제 및 헤미셀룰라아제 계열의 가수분해효소(효소A)와 아밀라아제(amylase) 계열의 가수분해효소(효소B)를 사용하였고, 상기 효소를 첨가 후 60℃에서 1시간 반응시키고, 고온에서 30분간 멸균처리하여 재료 각각의 배양배지화를 수행하였다. Each of turmeric, rice bran, and acidophilus, which removed foreign and fungal contamination, was subjected to enzymatic treatment and heat treatment sterilization for liquid culture culture. The enzymes used were hydrolytic enzymes (enzyme A) and amylase hydrolytic enzymes (enzyme B) of cellulase and hemicellulase system capable of degrading bark, muscle, roots and leaves. After the addition, the reaction was carried out at 60 DEG C for 1 hour and sterilized at a high temperature for 30 minutes to carry out culture medium culture of each material.
이후, 상기 배양배지에 별도로 배양한 담자균류균사를 첨가하였다. 담자균류균사 종균배양액을 10%(v/v) 접종하고 28~30℃ 및 pH 4.5~7의 조건에서 잔류 탄소원의 농도가 일정농도 이하로 고갈되는 시점에서 농축된 액상강황배지, 액상미강배지, 또는 액상산초배지를 첨가하는 유가식 공정으로 7~10일간 배양하였다. 상기의 담자균류균사는 구체적으로 약용 및 식용 가능한 담자균류인 상황버섯, 차가버섯, 표고버섯, 잎새버섯, 목이버섯, 눈꽃송이버섯, 치마버섯 등의 균사에서 어느 하나를 선택하여 사용할 수 있으며, 이들 담자균류 균사는 한국미생물보존센터(KCCM), 한국생명공학연구원 생명자원센터(KCTC), 한국세포주은행(KCLB), 한국농업미생물자원센터(KACC), 미국표준균주배양수록보존소(ATCC) 등의 기관에서 균주를 분양 받아 사용할 수 있다.Then, the mycelium of mycelia cultivated separately in the culture medium was added. When the concentration of the residual carbon source is depleted to a certain concentration or less under the conditions of 28 to 30 ° C and pH 4.5 to 7, inoculated with a 10% (v / v) culture of mycelia of mycelia, a concentrated liquid turmeric medium, Or an oil-in-water process in which liquid soy sauce medium was added for 7 to 10 days. The above mycelium of mycelia may be selected from mycelia such as mushroom, chaga mushroom, shiitake mushroom, mushroom mushroom, mushroom mushroom, snow mushroom mushroom, and skirting mushroom, which are medicinal and edible bacillus species, Ryu Myung-sa is a member of the Korean Society for Microbiology (KCCM), Korea Biotechnology Research Institute (KCTC), Korea Cell Line Bank (KCLB), Korea Agricultural Microbiology Resource Center (KACC) It is possible to use the strain in an institution.
상기 발효물에 대한 생물전환공정의 효소처리는 열수추출 후 섬유소분해효소 즉, 셀룰라아제(cellulase), 헤미셀룰라아제(hemicellulase), 펙티나아제(pectinase), 글루카나제(glucanase) 등의 다양한 효소제를 적당량(각 효소제의 제품설명서에서 제시하는 최적량)을 첨가하여 실시하였다. 효소로는 상업적으로 판매되고 있는 효소제 즉, 섬유소분해효소인 비스코자임(Viscozyme:Aspergillus 유래 복합물), 셀룰라아제(Cellulase: Aspergillus niger 유래), 필트라제(Filtrase:Tricoderma reesei 유래 복합물), 라피다제(Rapidase:Aspergillus niger 유래 복합물) 및 스미자임(Sumizyme:Aspergillus niger 유래 펙티나아제를 함유한 복합물)을 각 효소제의 제품설명서에서 제시하는 추천 농도(10% v/v)로 첨가하고, 50~60℃조건에서 1~3시간 동안 250 rpm으로 회전시키며 효소/기질반응을 수행하였다. 상기 발효 및 효소처리 생물전환공정에 의해 생산된 생물전환산물은 90℃, 1시간 효소 불활성화 과정 및 살균과정을 거친 후 동결건조하여 분말화 하였다. 상기의 강황, 미강, 또는 산초의 배양배지화, 생물전환공정 및 생물전환산물 분말 수득 과정을 도 1에 기재하였다.
The enzymatic treatment of the biotransformation process for the above fermented product is carried out by adding various enzymes such as cellulose decomposing enzyme, cellulase, hemicellulase, pectinase and glucanase after hot water extraction to an appropriate amount (The optimum amount indicated in the product manual of each enzyme preparation). Examples of the enzymes include commercially available enzymes such as a cellulase (a compound derived from Aspergillus niger), a filtrate (a compound derived from Filtrase (Tricoderma reesei)), a rapidase (a compound derived from a fibrinolytic enzyme, Viscozyme (Complex containing Aspergillus niger) and Sumizyme (a complex containing Aspergillus niger-derived pectinase) was added at the recommended concentration (10% v / v) given in the product manual of each enzyme preparation, The enzyme / substrate reaction was performed by rotating at 250 rpm for 1 to 3 hours. The bioconversion products produced by the fermentation and enzyme-treated bioconversion process were lyophilized and pulverized after enzyme inactivation and sterilization at 90 ° C for 1 hour. The culture medium, biotransformation process and biotransformation product powder of the above-described turmeric, rice bran, or sancho cultivation are shown in Fig.
실시예Example 2. 대식세포에서의 살모넬라 2. Salmonella in macrophages 탐식능Pickling 및 탐식 후 And after digestion 살해능Kill ability 향상 효능 평가 Evaluation of improvement efficacy
대식세포의 살모넬라 탐식능 및 살해능을 확인하기 위한 RAW264.7 세포주(마우스 대식세포주)를 세계세포주은행(ATCC)에서 구입하였고, 37℃, 5% CO2, 포화습도의 조건으로 15%의 보체 비활성화 FBS(heat-inactivated fetal bovine serum)와 100 유니트(units) 페니실린, 100 ㎍/㎖ 스트렙토마이신이 포함된 RPMI-1640 배지에서 배양하였다. RAW264.7 cell line (mouse macrophage line) was purchased from the World Cell Line Bank (ATCC) to confirm Salmonella robbing and killing ability of macrophages and 15% complement inactivated under conditions of 37 ° C, 5% CO 2 , The cells were cultured in RPMI-1640 medium containing heat-inactivated fetal bovine serum (FBS), 100 units of penicillin, and 100 μg / ml of streptomycin.
먼저, 대식세포의 살모넬라 탐식능 시험을 위해, 상기 RAW264.7 세포를 96 웰 배양접시에 1 x 104 세포/웰 농도로 접종한 후 하룻밤 배양하고, 담자균류균사 발효액을 4시간 처리하고 배지로 1회 세척하였다. 상기에 살모넬라 타이피뮤리움(salmonella typhimurium, 쥐티푸스균)을 1 x 106 CFU/㎖로 맞추어 10㎕ 처리하고(최종 1 x 104CFU), 30분 및 60분간 반응시킨 후 배지로 추가 세척하였다. 30㎍/㎖ 의 젠타마이신을 30분간 처리하여 배지 내 남아있는 살모넬라균을 제거하였고, PBS로 세척 후 트립신을 처리하여 세포를 수득한 후, 증류수 1㎖ (또는 0.1% triton X-100 in PBS 1㎖) 에 현탁하고 실온에서 30분간 교반하여 세포를 용해하였다. 이 후, 영양아가배지(nutrient agar)에 계열희석하여 접종하고, 16시간 배양하여 살모넬라균의 집락수를 측정하였다.First, RAW264.7 cells were inoculated in a 96-well culture dish at a concentration of 1 x 10 4 cells / well, and cultured overnight. For the salmonellosis test of macrophages, the mycelium fermented broth was treated for 4 hours, Lt; / RTI > Salmonella typhimurium (1 × 10 6 CFU / ml) was treated with 10 μl (final 1 × 10 4 CFU) and reacted for 30 minutes and 60 minutes, followed by further washing with a medium . The cells were treated with 1 ml of distilled water (or 0.1% triton X-100 in PBS 1), and the cells were washed with PBS and treated with trypsin. The cells were treated with 30 ml of gentamycin for 30 minutes to remove the remaining Salmonella in the medium. Ml) and the mixture was stirred at room temperature for 30 minutes to dissolve the cells. Subsequently, the cells were inoculated in series with a nutrient agar and cultured for 16 hours to determine the number of colonies of Salmonella.
대식세포의 살모넬라 탐식 후 살해능 시험을 위해서는 상기와 같은 방법으로 배양한 RAW264.7 세포를 96 웰 배양접시에 1 x 104 세포/웰 농도로 접종한 후 하룻밤 배양하고, 담자균류균사 발효액을 4시간 처리하고 배지로 1회 세척하였다. 상기에 살모넬라 타이피뮤리움(salmonella typhimurium, 쥐티푸스균)을 1 x 106CFU/㎖로 맞추어 10㎕ 처리하고(최종 1 x 104CFU), 1시간 반응시킨 후 배지로 추가 세척하였다. 30㎍/㎖ 의 젠타마이신을 2시간, 4시간 또는 8시간동안 처리하여 배지 내 남아있는 살모넬라균을 제거하였고, PBS로 세척 후 트립신을 처리하여 세포를 수득한 후, 증류수 1㎖ (또는 0.1% triton X-100 in PBS 1㎖) 에 현탁하고 실온에서 30분간 교반하여 세포를 용해하였다. 이 후, 영양아가배지(nutrient agar)에 계열희석하여 접종하고, 16시간 배양하여 살모넬라균의 집락수를 측정하였다.In order to test the viability after phagocytosis of macrophages, RAW264.7 cells cultured in the same manner as above were inoculated in a 96-well culture dish at a concentration of 1 x 10 4 cells / well and cultured overnight. Treated with time and washed once with medium. Salmonella typhimurium (1 × 10 6 CFU / ml) was treated with 10 μl (final 1 × 10 4 CFU) at 1 × 10 6 CFU / ml for 1 hour and further washed with the medium. The remaining Salmonella in the medium was removed by treatment with 30 μg / ml of gentamycin for 2 hours, 4 hours or 8 hours, washed with PBS, treated with trypsin to obtain cells, and then 1 ml of distilled water (or 0.1% triton X-100 in PBS (1 ml)) and the mixture was stirred at room temperature for 30 minutes to dissolve the cells. Subsequently, the cells were inoculated in series with a nutrient agar and cultured for 16 hours to determine the number of colonies of Salmonella.
실험 결과, 강황, 미강, 또는 산초의 모든 생물전환산물에서 처리 시간이 증가할수록 또는 처리 농도가 증가할수록 대식세포의 살모넬라 탐식능이 향상되는 것을 확인하였다. 대식세포의 살모넬라 탐식 후 살해능의 경우에는, 모든 생물전환산물에서 처리 2시간째에는 대조군(생물전환산물이 처리되지 않은 그룹)에 비하여 처리된 생물전환산물의 농도가 증가할수록 살모넬라 집락수가 많아 살모넬라 탐식능이 향상되었음을 확인할 수 있었으며, 4시간 및 8시간에서는 처리된 생물전환산물의 농도가 증가할수록 살모넬라 집락수가 급격히 감소하여 대조군보다 낮은 수치를 나타내어 살모넬라 탐식 후 살해능이 향상되는 것을 확인할 수 있었다. 즉, 생물전환산물 처리에 의해 대식세포의 살모넬라 탐식능과 함께 탐식 후 살해능이 향상되는 것을 확인할 수 있었다. 상기 결과를 도 2에 나타내었다. As a result, it was confirmed that the salmonellosis of macrophages was improved as the treatment time was increased or the treatment concentration was increased in all bioconversion products of turmeric, rice bran, or sancho. In the case of salmonellosis after macrophage salmonellosis, as the concentration of treated bioconverted products increased, the number of colonies of Salmonella increased at 2 hours after treatment in all bioconversion products compared to the control (group not treated with bioconversion products) In 4 hours and 8 hours, the salmonella colonies decreased rapidly as the concentration of treated bioconverted products increased. As a result, the numbers of Salmonella colonies were lower than those of the control group. In other words, it was confirmed that the biodegradation product treatment improves the kill ability after phagocytosis in addition to the salmonella attack ability of macrophages. The results are shown in Fig.
상기의 RAW264.7 세포주에 대한민국 진도산 또는 인도산 강황의 원물 또는 생물전환산물을 처리한 실험에서는, 대한민국 진도산과 인도산 모두 원물보다 생물전환산물에서 처리 농도가 증가할수록 대식세포의 살모넬라 탐식능이 크게 향상되었다. 특히, 10 ㎍/㎖ 이상의 생물전환산물이 처리되었을 때 대식세포의 탐식능이 대조군에 비하여 2배 이상 증가하는 것으로 나타났다. 대식세포의 살모넬라 탐식 후 살해능 효능 평가에서도 10 ㎍/㎖ 이상의 생물전환산물을 4시간 이상 처리하였을 때, 강황의 원산지에 상관없이 살모넬라 집락수가 크게 감소하는 것으로 나타났다. 상기의 강황 원산지별 원물 또는 생물전환산물의 대식세포 살모넬라 탐식능 및 살모넬라 살해능 결과를 도 3 및 도 4에 각각 나타내었다.
In the experiment in which RAW264.7 cell line was treated with Jindo jinhodang or Ganoderma lucidum, the colonization efficiency of macrophage Salmonella largely improved as the concentration of the bioconversion product increased in both Jindo and India . Especially, when macrophages were treated with 10 ㎍ / ㎖ or more of bioconversion products, the phagocytosis of macrophages was more than 2 times higher than that of the control. In the evaluation of viability after salmonellosis of macrophages, salmonella colonies were significantly decreased regardless of the origin of turmeric when the bioconversion product of 10 ㎍ / ㎖ or more was treated for 4 hours or more. Fig. 3 and Fig. 4 show the results of the macrophage salmonellosis and the salmonella kill ability of the raw or biotransformed product of each turmeric origin.
실시예Example 3. 치사량 살모넬라 감염 마우스 모델에서의 체내(in 3. Lethal dose Salmonella-infected mouse model (in vivovivo ) 효능 평가) Efficacy evaluation
마우스에 강황, 미강, 또는 산초의 생물전환산물을 10 mg/kg의 농도로 복강주사 또는 경구투여하였다. 복강주사의 경우, 1 ㎖ 주사기를 이용하여 10 mg/kg 농도에 맞게 200 ㎕ 복강주사하였고, 경구투여의 경우에는 10 mg/kg 농도로 사료에 섞어 식이로 섭취하게 하는 방법으로 투여하였다. 이 후, 치사량의 살모넬라균(1 x 105 CFU)을 주사하여, 치사량 살모넬라 감염 마우스 모델을 유도하였다. 상기 치사량 살모넬라 감염 모델 마우스의 생존율을 30일까지 관찰한 결과, 복강투여(도 5) 및 경구투여(도 6) 모두 대조군(생물전환산물이 처리되지 않은 그룹) 마우스가 6일 이내에 모두 사망한 데 비하여 생물전환산물이 처리된 그룹의 마우스는 생존기간이 월등히 긴 것을 확인하였다. 생물전환산물별로는 강황, 미강, 산초의 순서로 효능이 좋은 것으로 나타났다. Biotransformation products of turmeric, rice bran, or sancho were administered to mice by intraperitoneal injection or oral administration at a concentration of 10 mg / kg. In the case of intraperitoneal injection, 200 ㎕ of intraperitoneal injection was made at a dose of 10 mg / kg using a 1 ㎖ syringe and 10 mg / kg of the compound was orally administered to the diets. Later, a lethal dose of Salmonella (1 x 10 5 CFU) was injected to induce a mouse model of lethal Salmonella infection. The survival rate of the lethal Salmonella-infected model mouse was observed until 30 days. As a result, all of the mice in the control group (the group not treated with the biotransformation products) died within 6 days in both the peritoneal administration (FIG. 5) and the oral administration (FIG. 6) In contrast, the survival time of mice in the group treated with the bioconversion product was much longer. According to the biotransformation products, it was shown to be effective in order of turmeric, rice gruel and sancho.
상기의 복강투여 및 경구투여를 종합하여 비교하면, 전체적으로 생물전환산물을 경구투여의 경우보다 복강투여한 경우 치사량 살모넬라 감염 모델 마우스의 생존 시간이 증가하는 것을 확인하였다. 상기 결과를 도 7에 나타내었다.
Compared with the above-mentioned peritoneal administration and oral administration, overall survival time of the lethal Salmonella-infected model mouse was increased when the bioconversion product was administered to the abdominal cavity more than in the case of oral administration. The results are shown in FIG.
실시예Example
4. 4.
아치사량Abalone
살모넬라 감염 마우스 모델에서의 체내(in In a Salmonella-Infected Mouse Model (in
vivovivo
) 효능 평가) Efficacy evaluation
실시예 4-1. 아치사량 살모넬라 감염 마우스 모델 제작Example 4-1. Mouse model of infection with saliva Salmonella
마우스에 강황, 미강, 또는 산초의 생물전환산물을 10 mg/kg의 농도로 복강주사 또는 경구투여하였다. 복강주사의 경우, 1 ㎖ 주사기를 이용하여 10 mg/kg 농도에 맞게 200 ㎕ 복강주사하였고, 경구투여의 경우에는 10 mg/kg 농도로 사료에 섞어 식이로 섭취하게 하는 방법으로 투여하였다. 이 후, 아치사량의 살모넬라균(1 x 104 CFU)을 주사하여, 아치사량 살모넬라 감염 마우스 모델을 유도하였다.
Biotransformation products of turmeric, rice bran, or sancho were administered to mice by intraperitoneal injection or oral administration at a concentration of 10 mg / kg. In the case of intraperitoneal injection, 200 ㎕ of intraperitoneal injection was made at a dose of 10 mg / kg using a 1 ㎖ syringe and 10 mg / kg of the compound was orally administered to the diets. Subsequently, saline (1 x 10 4 CFU) of saliva was injected to induce mouse model of saliva Salmonella infection.
실시예 4-2. 아치사량 살모넬라 감염 마우스 복강 내 살모넬라 집락수 계측Example 4-2. Measurement of Salmonella colony count in the abdominal cavity of mice infected with saliva Salmonella infection
실시예 4-1의 방법으로 제조된 아치사량 살모넬라 감염 마우스 모델에 살모넬라 감염 48시간 후 마우스를 희생하여 복막을 드러낸 후, 10 ㎖ 의 PBS를 주입하고 복강 마사지 후 다시 PBS를 회수하였다. 회수된 PBS는 3500 rpm에서 20분간 원심분리 하였고, 새로운 PBS로 2회 세척 후, PBS 1 ㎖ 에 재부유하여 미생물용 고체배지(LB plate)에 접종하였다. 이를 24시간 배양 후, 살모넬라 집락수를 계측하였다. 실험 결과, 강황, 미강, 또는 산초의 모든 생물전환산물 처리 그룹에서 대조군(생물전환산물이 처리되지 않은 그룹)에 비하여 월등히 감소된 살모넬라 집락수 결과가 확인되었다. 생물전환산물 투여방법에 따른 비교에서는 모든 생물전환산물 처리 그룹에서 경구투여의 경우보다 복강투여한 경우 살모넬라 집락수가 더 많이 감소하는 것으로 나타났다. 상기 결과를 도 8에 나타내었다.
Mice were sacrificed 48 hours after the Salmonella infection, and the peritoneum was sacrificed. Then, 10 ml of PBS was injected, and PBS was recovered after peritoneal massage. The recovered PBS was centrifuged at 3500 rpm for 20 minutes, washed twice with fresh PBS, resuspended in 1 ml of PBS, and inoculated on a solid medium (LB plate) for microorganism. After culturing for 24 hours, the number of Salmonella colonies was counted. The results showed that the Salmonella colony counts were significantly reduced in all the bioconversion product treatment groups of turmeric, rice bran, or sancho, as compared to the control (the group not treated with bioconversion product). Comparisons by bioconversion product dosing showed that Salmonella colonies decreased more in all bioconversion product groups when administered intraperitoneally than oral doses. The results are shown in FIG.
실시예 4-3. 아치사량 살모넬라 감염 마우스 비장 내 림프구수 계측Example 4-3. Measuring the number of lymphocytes in mouse spleen salmonella-infected mice
실시예 4-1의 방법으로 제조된 아치사량 살모넬라 감염 마우스 모델에 살모넬라 감염 48시간 후 마우스를 희생하여 비장을 적출하고 스테인레스강 메시(stainless steel mesh)를 이용하여 전체 비장세포(total splenocyte)를 분리하였다. 분리된 비장세포는 2 x 105 cells/well 의 농도로 맞추어 96-웰 배양접시에 접종하였고, 0.5 ㎍/㎖의 콘카나발림 A(Concanavalin A)와 1 ㎍/㎖의 LPS를 처리하여 림프구의 증식을 유도하였다. 72시간 후 MTT 어세이를 통해 세포 수를 측정하였으며, 살모넬라에 감염되지 않은 건강한 마우스의 전체 비장세포 수를 1로 하여 증가량을 계산하였다. 실험 결과, 대조군(생물전환산물이 처리되지 않은 그룹) 마우스의 비장 림프구가 건강한 마우스에 비하여 약 1.5배 증가한 것에 비하여, 강황, 미강, 또는 산초의 모든 생물전환산물 처리 그룹에서 마우스의 비장 림프구가 건강한 마우스에 비하여 약 2배 증가한 것을 확인하였다. 특히 생물전환산물을 복강투여한 경우에는 경구투여한 경우보다 마우스의 비장 림프구가 더 많이 증가하였다. 상기 결과를 도 9에 나타내었다.
Mice were sacrificed 48 hours after the salmonella infection in a mouse model of saliva Salmonella infected by the method of Example 4-1, and the total spleen cells (total splenocytes) were isolated using a stainless steel mesh Respectively. Separated spleen cells were inoculated into 96-well culture dishes at a concentration of 2 x 10 5 cells / well, treated with 0.5 μg / ml of Concanavalin A and 1 μg / ml of LPS, Proliferation was induced. After 72 hours, the number of cells was measured using an MTT assay, and the amount of increase was calculated by setting the total number of splenocytes in healthy mice not infected with Salmonella to 1. Experimental results showed that the spleen lymphocytes of the mice in the control group (the group not treated with the biotransformation product) increased about 1.5 times as compared with the healthy mice, Mice were approximately 2-fold increased compared to mice. Especially, when the bioconversion product was administered intraperitoneally, the number of mouse spleen lymphocytes was increased more than that of oral administration. The results are shown in Fig.
실시예 4-4. 아치사량 살모넬라 감염 마우스 비장 내 사이토카인 측정Example 4-4. Measuring cytokine in mouse spleen infected with saliva Salmonella infection
실시예 4-1의 방법으로 제조된 아치사량 살모넬라 감염 마우스 모델에 살모넬라 감염 48시간 후 마우스를 희생하여 비장을 적출하고, 적출한 비장을 균질기(homogenizer)로 분쇄한 후, ELISA 방법을 이용하여 염증반응과 관련이 있는 사이토카인인 IL-1β(R&D system, MLB00C), Th1 면역반응과 관련된 사이토카인인 IFN-γ(R&D system, MIF00), IL-2(R&D system, M2000) 및 Th2 면역반응과 관련된 사이토카인인 IL-5(R&D system, M5000)의 사이토카인 수치를 측정하였다. 상기 사이토카인의 측정은 제조사의 권장 프로토컬에 따라 시행하였다. 실험 결과, 염증반응과 관련이 있는 사이토카인인 IL-1β와 함께 Th1 면역반응과 관련된 사이토카인인 IFN-γ 및 IL-2에서, 강황, 미강, 또는 산초의 모든 생물전환산물 처리 그룹에서 대조군(생물전환산물이 처리되지 않은 그룹)에 비하여 사이토카인 수치가 증가한 것을 확인하였다. 또한 전반적으로, 생물전환산물을 복강투여한 경우에 경구투여한 경우보다 사이토카인 수치가 더 많이 증가하였다. 그러나 IL-5 의 경우에는 생물전환산물 처리 그룹과 대조군(생물전환산물이 처리되지 않은 그룹) 사이에 큰 차이가 없는 것으로 나타났다. 상기 결과를 도 10에 나타내었다.Mice were sacrificed 48 hours after infection with Salmonella and the spleen was sacrificed. The spleen was pulverized with a homogenizer, and then subjected to ELISA IL-2 (R & D system, M2000) and Th2 immunoreactivity, which are related to the inflammatory response, IL-1β (R & D system, MLB00C), the cytokine IFN-γ And cytokine levels of IL-5 (R & D system, M5000), a cytokine associated with IL-5. Measurement of the cytokine was carried out according to the manufacturer's recommended protocol. As a result, the IFN-γ and IL-2, which are cytokines related to the Th1 immune response together with IL-1β, which is a cytokine related to the inflammatory response, were observed in all the bioconversion product treatment groups of turmeric, Compared with the control group (group not treated with the bioconversion product). Overall, the cytokine levels were also increased more when peritoneal administration of the bioconversion product was administered than by oral administration. However, in the case of IL-5, there was no significant difference between the bioconversion product treatment group and the control group (the untreated bioconversion product group). The results are shown in FIG.
아치사량 살모넬라 감염 마우스 모델에 강황생물전환산물을 처리한 실험군에서 상기의 IL-1β, IFN-γ, IL-2 및 IL-5 에 추가로 IL-4(R&D system, M4000B), IL-6(R&D system, M6000B), IL-10(R&D system, M1000) 및 IL-12(R&D system, M1270) 사이토카인의 수치를 측정하였다. 추가된 사이토카인들의 측정은 상기의 IFN-γ, IL-1β, IL-2 및 IL-5와 마찬가지로 제조사의 권장 프로토컬에 따라 시행하였다. 실험 결과, 염증반응과 관련이 있는 사이토카인인 IL-1β, IL-6와 함께 Th1 면역반응과 관련된 사이토카인인 IFN-γ, IL-2, IL-12의 발현량은 살모넬라 감염 마우스에 강황 또는 미강의 생물전환산물이 복강 또는 경구로 투여된 그룹에서 살모넬라 감염 마우스 대조군에 비하여 증가되었으나, Th2 면역반응과 관련된 사이토카인인 IL-4, IL-5 와 함께 Treg 면역반응과 관련된 사이토카인인 IL-10은 살모넬라 감염 마우스 대조군과 비교하여 유의한 차이가 없는 것으로 나타났다. 즉, 살모넬라 감염 후 일어난 면역반응에 있어서 강황 또는 미강의 생물전환산물을 투여한 경우 Th2 면역반응과 관련된 사이토카인의 생성량은 변화가 거의 없는 반면, Th1 면역반응과 관련된 사이토카인의 생성량은 증가시킴을 확인한 바, 이는 Th1 면역반응이 바이러스와 세포내 감염 미생물(박테리아, 원생동물)의 방어와 관련된 세포성 면역에 중요하고 또한 살모넬라가 대식세포내에 감염을 일으키는 세포내 감염 미생물임을 고려할 때 상기 강황, 미강, 또는 산초의 모든 생물전환산물은 면역학적 근거에 기초해 효과적으로 대처할 수 있을 것으로 기대된다. 강황 또는 미강의 상기 결과를 도 11 및 도 12에 각각 나타내었다.
In addition to IL-1β, IFN-γ, IL-2 and IL-5, IL-4 (R & D system, M4000B), IL-6 (R & D system, M12000), IL-10 (R & D system, M1000) and IL-12 (R & D system, M1270) The measurement of added cytokines was carried out according to the manufacturer's recommended protocol as with IFN-y, IL-1 beta, IL-2 and IL-5. The expression levels of IFN-γ, IL-2, and IL-12, which are cytokines related to the Th1 immune response in combination with the IL-1β and IL-6 cytokines related to the inflammatory response, In addition, IL-4 and IL-5, which are cytokines related to Th2 immune response, and IL-5, which are related to Treg immunoreactivity, were significantly increased in IL- 10 was not significantly different from the control group of Salmonella infected mice. In other words, the cytokine production associated with the Th2 immune response was not changed when the biotransformation product of turmeric or rice bacilli was administered in the immune response after the salmonella infection, but the amount of the cytokine related to the Th1 immune response was increased Considering that Th1 immune response is important for cellular immunity related to the defense of viruses and intracellular infecting microorganisms (bacteria, protozoa), and also because Salmonella is an intracellular infecting microorganism causing infection in macrophages, , Or all bioconversion products of Sancho are expected to be able to effectively cope based on immunological evidence. The above results of turmeric or corona are shown in Fig. 11 and Fig. 12, respectively.
실시예 4-5. 아치사량 살모넬라 감염 마우스의 간 손상 억제 효과 확인Example 4-5. Identification of liver injury inhibition effect of mice infected with saliva Salmonella
살모넬라 감염으로 인해 발생하는 전신성 염증반응에 의한 조직의 손상을 면역조절소재의 투여에 의해 억제할 수 있는지 확인하기 위하여 10mg/kg의 강황(생물전환)소재를 2주간 식이를 통하여 투여하였고, 1 x 10⁴CFU의 ATCC14028 살모넬라 균주를 복강 내로 주사하여 감염을 유도하였다. 감염 2일 후 전신 염증반응이 유도된 마우스를 희생하여 간 조직을 분리 후, 염색하여 관찰한 결과 강황(생물전환)소재를 투여한 마우스에서는 대조군에 비해 간 조직의 손상이 억제된 것이 확인되었다. 상기 결과를 도 13에 나타내었다.
To determine if tissue damage caused by the systemic inflammatory response caused by Salmonella infection could be inhibited by the administration of an immunomodulatory agent, 10 mg / kg of turmeric (bioconversion) material was administered via diet for 2 weeks and 1 x Infection was induced by intraperitoneal injection of ATCC14028 salmonella strain of 10⁴ CFU. At 2 days after the infection, liver tissue was isolated and sacrificed at the expense of systemic inflammatory response, and it was observed that the liver tissue damage was suppressed in the mice treated with the carbohydrate (bioconversion) material compared with the control. The results are shown in Fig.
실시예 5. LPS 유도 패혈증 모델에서의 체내(in vivo) 효능 평가Example 5. Evaluation of in vivo efficacy in LPS-induced sepsis model
실시예 5-1. 생물전환산물 투여량에 따른 효능 평가Example 5-1. Assessment of efficacy by bio-conversion product dose
마우스에 강황, 미강, 또는 산초의 생물전환산물을 1, 10 및 100 mg/kg의 농도로 1 ㎖ 주사기를 이용하여 2주간 200 ㎕ 복강주사로 투여하고 치사량의 LPS (20 ㎍/kg)와 갈락토사민(galactosamine, 700 mg/kg)을 주사하여 패혈증을 유도하였다. 상기 패혈증 모델 마우스의 시간별 생존율을 60시간까지 관찰하였다. 실험 결과, 강황, 미강, 또는 산초의 모든 생물전환산물에서 처리된 생물전환산물의 농도가 증가할수록 패혈증 모델 마우스의 생존시간이 증가하는 것을 확인하였다. 상기 결과를 도 14에 나타내었다.
Biotransformation products of turmeric, rice gruel, or sancho were administered to mice in a dose of 1, 10, and 100 mg / kg by intraperitoneal injection of 200 ㎕ for 2 weeks using a 1 ㎖ syringe, and lethal dose of LPS (20 ㎍ / kg) Sepsis was induced by injecting galactosamine (700 mg / kg). The survival rate over time of the sepsis model mice was observed for up to 60 hours. Experimental results showed that the survival time of the sepsis model mice was increased as the concentration of bioconverted products treated in all bioconversion products of turmeric, rice bran, or sancho was increased. The results are shown in Fig.
실시예 5-2. 생물전환산물 투여경로에 따른 효능 평가Example 5-2. Evaluation of efficacy according to route of biotransformation product
마우스에 강황, 미강, 또는 산초의 생물전환산물을 10 mg/kg의 농도로 복강주사 또는 경구투여하였다. 복강주사의 경우, 1 ㎖ 주사기를 이용하여 10 mg/kg 농도에 맞게 200 ㎕ 복강주사하였고, 경구투여의 경우에는 10 mg/kg 농도로 사료에 섞어 식이로 섭취하게 하는 방법으로 투여하였다. 이 후, 치사량의 LPS (20 ㎍/kg)와 갈락토사민(galactosamine, 700 mg/kg)을 주사하여 패혈증을 유도하고, 상기 패혈증 모델 마우스의 시간별 생존율을 60시간까지 관찰하였다. 실험 결과, 강황, 미강, 또는 산초의 모든 생물전환산물에서 경구투여의 경우보다 복강투여한 경우 패혈증 모델 마우스의 생존시간이 증가하는 것을 확인하였다. 또한, 투여방법에 상관없이, 투여군 모두에서 대조군(생물전환산물이 처리되지 않은 그룹)에 비하여 생존시간이 증가하였다. 상기 결과를 도 15에 나타내었다.Biotransformation products of turmeric, rice bran, or sancho were administered to mice by intraperitoneal injection or oral administration at a concentration of 10 mg / kg. In the case of intraperitoneal injection, 200 ㎕ of intraperitoneal injection was made at a dose of 10 mg / kg using a 1 ㎖ syringe and 10 mg / kg of the compound was orally administered to the diets. Thereafter, sepsis was induced by injecting LPS (20 / / kg) and galactosamine (700 mg / kg) at a lethal dose and the survival rate over time of the sepsis model mice was observed for 60 hours. Experimental results showed that the survival time of septicemic mice was increased in all bioconversion products of turmeric, rice bran, and sancho. In addition, regardless of the method of administration, the survival time was increased in the administration group as compared to the control (the group in which the biotransformation product was not treated). The results are shown in Fig.
상기의 실험 결과를 동일 투여방법 기준 생물전환산물별로 비교하였다. 복강투여와 경구투여 모두 동일한 10 mg/kg 투여의 경우, 강황, 미강, 산초의 순서로 효능이 좋은 것으로 나타났으며, 대조군이 가장 낮은 효과를 나타내었다. 상기 결과를 도 16에 나타내었다.
The results of the above experiments were compared according to the biological conversion products of the same administration method. In the case of 10 mg / kg administration of both peritoneal administration and oral administration, the efficacy was shown to be in the order of turmeric, rice gruel, and sancho, and the control group had the lowest effect. The results are shown in Fig.
실시예Example 6. 6. 미강과Rice bran 흑미강의Black rice lecture 면역자극효과 비교 Comparison of immunostimulatory effects
미강은 크게 백미강과 흑미강으로 구분된다. 본 발명에서의 미강은 현미를 백미로 도정하는 공정에서 분리되는 하얀 빛깔의 쌀겨를 의미하는 것으로, 흑미강과 대조될 수 있다.Rice cores are divided into white rice and black rice. In the present invention, rice bran means rice bran of white color separated in the process of converting brown rice into white rice, which can be contrasted with black rice.
미강과 흑미강의 생물전환산물에 있어서 면역자극효과를 비교하기 위하여, 5% DMEM 배지를 사용하여 마크로파지(대식세포)의 배양 및 NO(nitric oxide) 분석을 수행하였다. 대식세포(마크로파지)에 대한 면역활성 측정을 위해 대식세포는 세포주를 사용하거나 또는 마우스 복강에 5 ㎖의 1X PBS를 주입하고 마우스를 충분히 흔들어 준 다음, 복강내 대식세포를 주사기를 이용하여 수득한 마우스 복강 대식세포를 사용하여 96 웰 배양접시에 웰당 1 x 106 cell/㎖의 농도로 0.2 ㎖씩 세포를 계대배양 하였다. 24시간 배양 후 현미경으로 세포의 배양상태를 확인한 뒤, 배지를 완전히 제거한 다음 혈청이 첨가되지 않은 배지로 2~3회 세척하였다. 이후, 각 웰에 미강 또는 흑미강 생물전환산물 시료가 첨가된 5% FBS 배지를 0.5 ㎖씩 채워주고, 시료의 농도가 1/64, 1/32, 1/16, 1/8, 1/4, 1/2, 1, 2, 4, 8, 및 16 ㎍/㎖이 되도록 희석하였다. 또한, 양성대조군으로 최종 농도를 0.1㎍/㎖의 LPS (200㎍/mL stock)를 처리하였다. 시료가 처리된 24 웰 배양접시를 20 내지 24시간 배양 후, 각 웰의 배양액을 100㎕씩 취하여 원심분리(12,000 rpm, 5분)하여 상등액을 취하여 NO 분석에 사용하였다. In order to compare the immunostimulatory effect of the bioconversion products of rice bran and black rice, culture of macrophages (macrophages) and NO (nitric oxide) analysis were carried out using 5% DMEM medium. For measurement of immunological activity against macrophages (macrophages), macrophages were used either as a cell line or 5 ml of 1X PBS was injected into mouse abdominal cavity, the mice were sufficiently shaken, and then intraperitoneal macrophages were infected with mouse Cells were subcultured in a 96-well culture dish using peritoneal macrophages at a concentration of 1 x 10 6 cells / ml per well in 0.2 ml portions. After culturing for 24 hours, the culture conditions of the cells were checked with a microscope, and the medium was completely removed, followed by washing with a medium containing no serum for 2-3 times. Then, each well was filled with 0.5 ml of 5% FBS medium containing a sample of rice bran or black rice biotransformation product, and the concentration of the sample was 1/64, 1/32, 1/16, 1/8, 1/4, 1/2, 1, 2, 4, 8, and 16 [mu] g / ml. In addition, the final concentration was treated with 0.1 μg / ml of LPS (200 μg / ml stock) as a positive control. After culturing the 24-well culture dishes treated with the samples for 20 to 24 hours, 100 μl of the culture solution of each well was centrifuged (12,000 rpm, 5 minutes), and the supernatant was taken for NO analysis.
상기의 방법으로 준비된 시료를 96 웰 배양접시의 각 웰에 넣고, 그리스 용액(griess reagent, ① N-(1-naphtyl) ethylene diamine dihydrochloride ②sulfanilamide)을 100 ㎕ 분주한 후 1분간 반응시켰다. 이후, ELISA 리더기를 이용하여 540 내지 550 nm의 파장에서 흡광도 값을 측정하여 다양한 원물 및 생물전환발효산물의 면역활성 역가를 측정하였다. 그 결과를 도 17에 기재하였다.The prepared samples were put into each well of a 96-well culture dish and 100 μl of a grease solution (① N- (1-naphtyl) ethylene diamine dihydrochloride ②sulfanilamide) was dispensed and reacted for 1 minute. Then, the absorbance values were measured at a wavelength of 540 to 550 nm using an ELISA reader, and the immobilized activity titers of various raw materials and bioconversion fermentation products were measured. The results are shown in Fig.
실험 결과, 시료 처리 3일째에 미강 생물전환산물이 처리된 시료는 1 ㎍/㎖ 시료의 농도에서 NO 포화를 나타낸 반면에 흑미강 생물전환산물이 처리된 시료는 4 ㎍/㎖ 시료의 농도에서 NO 포화를 나타내었다. 또란 시료 처리 5일째에는 미강 생물전환산물이 처리된 시료는 1 ㎍/㎖ 시료의 농도에서 NO 포화를 나타낸 반면에 흑미강 생물전환산물이 처리된 시료는 2 ㎍/㎖ 시료의 농도에서 NO 포화를 나타내었다. 이것으로 미강 생물전환산물이 흑미강 생물전환산물에 비하여 2배 이상의 면역자극효과가 있다는 것을 알 수 있었다.
As a result of the experiment, the samples treated with rice bran bioconversion product on the third day of the treatment showed NO saturation at the concentration of 1 ㎍ / ml, whereas the samples treated with the brown rice bioconversion product showed NO Saturation. On the 5th day after trough treatment, samples treated with rice bran bioconversion products showed NO saturation at the concentration of 1 ㎍ / ㎖ sample, whereas samples treated with brown rice bioconversion product showed NO saturation at the concentration of 2 ㎍ / ㎖ sample Respectively. This indicates that the rice bran bioconversion product has an immunostimulatory effect twice as much as that of the black rice bioconversion product.
Claims (13)
(b) 상기 (a)단계에서의 가수분해물에 담자균류균사를 접종하고, 배양하여 발효물을 제조하는 단계; 및
(c) 상기 (b)단계에서의 배양공정에 의해 생산된 발효물에 섬유소분해효소를 처리하는 단계를 포함하는, 미강생물전환산물의 제조방법.
(a) hydrolyzing rice bran with enzymes of the cellulase and hemicellulase family and amylase family;
(b) inoculating the hydrolyzate of step (a) with mycelia of mycelia and culturing to produce a fermented product; And
(c) treating the fermented product produced by the culturing step in the step (b) with a fibrinolytic enzyme.
상기 담자균류균사는 상황버섯, 차가버섯, 표고버섯, 잎새버섯, 목이버섯, 눈꽃송이버섯, 치마버섯으로 이루어진 그룹으로부터 선택된 어느 하나 이상인, 미강생물전환산물의 제조방법.
The method according to claim 1,
Wherein said mycelium of mycelia is at least one selected from the group consisting of mushroom, mushroom, shiitake, mushroom, mushroom, mushroom, and mushroom.
상기 (c)단계에서의 섬유소분해효소는 세포벽 구성성분을 분해할 수 있는 효소인 것인, 미강생물전환산물의 제조방법.
The method according to claim 1,
Wherein the cellulolytic enzyme in step (c) is an enzyme capable of degrading a cell wall component.
상기 섬유소분해효소는 셀룰라아제, 헤미셀룰라아제, 펙티나아제 및 글루카나제로 이루어진 그룹으로부터 선택된 어느 하나 이상인, 미강생물전환산물의 제조방법.
The method of claim 3,
Wherein the fibrinolytic enzyme is at least one selected from the group consisting of cellulase, hemicellulase, pectinase, and glucanase.
A pharmaceutical composition comprising, as an active ingredient, a rice bran biotransformation product obtained by hydrolyzing rice bran with an enzyme of the cellulase and hemicellulase family enzymes and an amylase family enzyme and treating the fermented product cultured by inoculating mycelia with a mycelium,
상기 약학조성물은 살모넬라 감염의 예방 또는 치료 목적인, 약학조성물.
6. The method of claim 5,
The pharmaceutical composition is intended for the prevention or treatment of Salmonella infection.
상기 약학조성물은 살모넬라에 대한 대식세포의 탐식능 및 탐식 후 살해능을 향상시키는, 약학조성물.
The method according to claim 6,
Wherein said pharmaceutical composition enhances the phagocytosis and phagocytosis of macrophages against Salmonella.
상기 약학조성물은 살모넬라 감염시 Th1 도움 T세포(Th1 helper T cell) 면역반응 유도를 향상시키는, 약학조성물.
The method according to claim 6,
Wherein said pharmaceutical composition enhances Th1 helper T cell immune response induction upon Salmonella infection.
상기 약학조성물은 패혈증의 예방 또는 치료 목적인, 약학조성물.
6. The method of claim 5,
Wherein said pharmaceutical composition is for the prevention or treatment of sepsis.
A food composition comprising, as an active ingredient, a rice bran biotransformation product obtained by hydrolyzing rice bran with an enzyme of the cellulase and hemicellulase family enzymes and an amylase family enzyme and treating the fermented product cultured by inoculating mycelia with a mycelium of fungi,
A feed composition comprising a rice bran bioconversion product obtained by hydrolyzing rice bran with an enzyme of the cellulase and hemicellulase family enzymes and an amylase family enzyme and treating the fermented product cultivated by inoculating the mycelium of the bacillus family with a cellulose decomposing enzyme as an active ingredient.
상기 사료조성물은 소, 돼지, 가금류 등의 사육용인, 사료조성물.
12. The method of claim 11,
The feed composition is suitable for breeding cattle, pigs, poultry and the like.
상기 가금류는 닭, 오리, 칠면조, 거위, 화조, 집비둘기 및 메추리 등으로 구성된 군으로부터 선택된 어느 하나 이상인, 사료조성물.
13. The method of claim 12,
Wherein the poultry is any one or more selected from the group consisting of chickens, ducks, turkeys, geese, birds, pigeons and quails.
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