KR101425466B1 - Leaf of Smilax china with Aspergillus species, method for preparing the same and use of the same - Google Patents
Leaf of Smilax china with Aspergillus species, method for preparing the same and use of the same Download PDFInfo
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- KR101425466B1 KR101425466B1 KR1020130044584A KR20130044584A KR101425466B1 KR 101425466 B1 KR101425466 B1 KR 101425466B1 KR 1020130044584 A KR1020130044584 A KR 1020130044584A KR 20130044584 A KR20130044584 A KR 20130044584A KR 101425466 B1 KR101425466 B1 KR 101425466B1
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- South Korea
- Prior art keywords
- fermented
- leaf
- aspergillus
- fermentation
- extract
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Abstract
본 발명의 일 측면은 청미래덩굴 잎을 아스퍼질러스속 미생물로 발효시켜 얻은 산물인 발효 청미래덩굴 잎을 제공한다. 또한, 본 발명의 다른 측면은 청미래덩굴 잎을 아스퍼질러스속 미생물로 발효시켜 얻은 산물인 발효 청미래덩굴 잎으로부터 추출한 발효 청미래덩굴 잎 추출물을 제공한다. 본 발명에 따른 발효 청미래덩굴 잎은 아스퍼질러스속 미생물로 발효되어 발효 전보다 향상된 잔틴 옥시다아제(Xanthine oxidase) 억제 활성 또는 알데하이드 옥시다아제(aldehyde oxidase) 억제 활성을 나타낸다. 또한, 본 발명에 따른 발효 청미래덩굴 잎은 발효 전보다 향상된 관능적 특성을 나타낸다. 따라서, 본 발명에 따른 발효 청미래덩굴 잎으로 제조한 발효 차나 기능성 음료는 상업적 품질이 우수하고 시음자의 건강을 유지시키거나 개선하는데에 유리하다. 또한, 본 발명에 따른 발효 청미래덩굴 잎의 추출물 또는 이의 분획물은 통풍, 간 손상, 비만, 고지혈증, 지방간 등을 예방, 개선 또는 치료하기 위한 의약품 소재 또는 기능성 식품 소재로 매우 유용하다.One aspect of the present invention provides a fermented bluefin tuna leaf, which is a product obtained by fermenting a bluefin tuna vine leaf with an aspherical microorganism. In another aspect of the present invention, there is provided a fermented blueberry leaf vine leaf extract extracted from a fermented blueberry leaf vine leaf, which is a product obtained by fermenting a blueberry leaf blade with an aspergillus microorganism. According to the present invention, the fermented Cynthia latifolia leaf is fermented into an aspergillus microorganism and exhibits an activity of inhibiting xanthine oxidase or aldehyde oxidase, which is more improved than before fermentation. In addition, the fermented blueberry leaf of the present invention exhibits improved sensory properties before fermentation. Therefore, the fermented tea or the functional beverage produced from the fermented green tea leaf of the present invention has an excellent commercial quality and is advantageous for maintaining or improving the health of the tasting person. In addition, the fermented green tea leaf extract or its fraction according to the present invention is very useful as a pharmaceutical material or a functional food material for preventing, ameliorating or treating gout, liver damage, obesity, hyperlipemia, fatty liver and the like.
Description
본 발명은 청미래덩굴 잎으로부터 유래하는 유용한 기능성 소재 등에 관한 것으로서, 더 상세하게는 특정 미생물에 의해 발효된 청미래덩굴 잎, 이의 제조방법 및 이의 용도에 관한 것이다.The present invention relates to useful functional materials derived from Cheongmyeongvle leaf, and more particularly to a Cheongmyeongvle leaf fermented by a specific microorganism, a production method thereof, and a use thereof.
또한, 본 발명은 특정 미생물에 의해 발효된 청미래덩굴 잎의 추출물 내지 분획물, 이의 제조방법 및 이의 용도에 관한 것이다. Further, the present invention relates to an extract or a fraction of Cheongmyeongvle leaf fermented by a specific microorganism, a method for producing the same, and a use thereof.
경제개발로 인한 공업화와 더불어 식생활의 서구화로 공기, 수질을 포함한 환경오염 등 물리, 화학적 환경변화와 운동부족, 대사의 불균형, 각종 스트레스의 증가 등으로 다양한 생활 습관병이 급격히 증가하고 있다 (Carr 2003). 이 같은 병리적 상태에서는 체내에 반응성 산소종(reactive oxygen spices; 이하 ROS라 함) 생성계 효소인 XO(xanthine oxidase)와 AO(aldehyde oxidase) 활성이 증가하고(Halliwell 2006) 체내에 산화를 촉진하는 H2O2(hydrogen peroxide), O2 -(superoxide anion radical), 1O2(singlet oxygen), ·OH(hydroxyl radical) 등과 같은 반응성이 높은 반응성 산소종(reactive oxygen spices; 이하 ROS라 함)의 생성이 촉진 된다(Halliwell 2006). 이들 ROS는 생체막의 불포화지방산을 산화시켜 조직 내에 과산화지질을 축적시키고 조직의 산화적 손상을 초래하며 효소의 불활성화, 세포 노화, 동맥경화, 당뇨병, 뇌졸중 및 암과 같은 질병을 일으킨다 (Halliwell 2006). 생명체는 ROS의 공격을 방어하는 아스코브산(ascorbic acid), 글루타치온(glutathione), 셀레늄(selenium), 토코페롤(tocopherol), 리보플라빈(riboflavin) 등과 같은 비단백성 항산화 물질과 슈퍼옥사이드 디스푸타아제(superoxide dismutase), 카탈라아제(catalase), 글루타치온 페록시다아제(glutathione peroxidase), 글루타치온 S-전이효소(glutathione S-transferase, GST) 등과 같은 항산화계 효소들이 존재하여 ROS로 인한 독성을 경감시키며, 각종 질병의 예방과 노화를 지연시키는 것으로 알려져 있으나 (Jeon 등 2001; Moon 등 2001; De Haan 등 1995), 과잉의 산화적 스트레스에 의한 다양한 질병들로부터 생체를 보호하기 위하여 항산화 시스템을 강화시킬 수 있는 새로운 생리활성물질의 끊임없는 개발이 요구되고 있다.In addition to industrialization due to economic development, diverse lifestyle diseases have been increasing rapidly due to westernization of dietary habits due to physical and chemical environment changes such as air and water quality, lack of exercise, unbalanced metabolism, and various stresses (Carr 2003) . In this pathological state, the activity of xanthine oxidase (XO) and aldehyde oxidase (AO), which are reactive oxygen species (ROS) producing enzymes, increases in the body (Halliwell 2006) Reactive oxygen spices (hereinafter referred to as ROS), such as H 2 O 2 (hydrogen peroxide), O 2 - (superoxide anion radical), 1 O 2 (singlet oxygen) (Halliwell 2006). These ROSs oxidize unsaturated fatty acids in the biomembrane to accumulate lipid peroxides in tissues, causing oxidative damage of tissues and causing diseases such as inactivation of enzymes, cell senescence, arteriosclerosis, diabetes, stroke and cancer (Halliwell 2006) . The organism is composed of a nonspecific antioxidant such as ascorbic acid, glutathione, selenium, tocopherol, riboflavin and the like which protect against ROS attack and superoxide dismutase ), Catalase, glutathione peroxidase, glutathione S-transferase (GST), etc. are present to reduce the toxicity caused by ROS and to prevent various diseases (2001) and De Haan et al. (1995)), a new biologically active substance that can enhance the antioxidant system to protect the body from various diseases caused by excessive oxidative stress Is required to be continuously developed.
통풍은 관절염의 원인이 되는 대사성질환의 일종으로 요산(uric acid)의 과잉생성 또는 배설저하로 유발된 고요산 혈증으로 인하여 관절부위에 요산의 결정체가 축적되어 발생하는 염증으로 통증을 일으키는 질환이다(Chena 등 2011). 또한 요산은 푸린(purine)의 최종 대사산물로 XO(xanthine oxidase)에 의하여 잔틴(xanthine)으로부터 생성된다. XO는 체내 스트레스 등 병리적인 상태에서 크게 활성화하여 작용한다(Oei 등 1982; Urano 등 1991; Ham과 Kim 2004). 과요산증의 90%는 신장의 불충분한 배설작용에 의하여 일어나며 10%정도는 요산의 과도한 생성 때문에 일어난다(Wortmann 2002). 요산의 불충분한 배설은 신장내 요산염(urate) 수송계의 이상으로 일어나는 반면 과도한 요산은 식이 푸린(purine) 또는 XO의 활성증가로 생성된다(Anzai 등 2005; Caulfield 등 2008). 이에 따라 최근 항과요산증제제 또는 XO 저해제들이 많이 개발되고 있으나 소화기 장애, 골수생성억제, 신장장애, 과민성 증후군 등을 동반하는 등 부작용이 알려져 있다 (Horiuchi 등 2000; Zhu 등 2004; Robert와 Terkeltaub 2003). AO(aldehyde oxidase) XO와 분자량이 유사하고 플라빈 아데닌 디뉴클레오티드(flavin adenine dinucleotide, FAD), 몰리브덴(molybdenum) 및 철-유황과 같은 보결분자단을 가지고 있으며 생체 내 작용에 있어서 상호 밀접한 관련이 있으나 기질의 특이성에는 차이가 있는 것으로 보고되고 있다(Beedham 1987). AO는 간, 폐 및 신장 등의 세포질에 다량으로 존재하는 효소(Critchley 등 1992; Al-Salmy 2001)로 항암제(methotrexate)와 항바이러스제(famciclovir)와 같은 약물을 포함한 알데하이드(aldehyde) 및 질소함유 헤테로고리 화합물의 산화를 촉진하며(Beedham 1987; Clarke 등 1995; Jordan 등 1999; Kawashima 등 1999; Kitamura 등 1999), 전자공여체의 존재 하에서는 황산화물(sulfoxides), 질소산화물(N-oxides), 니트로사민(nitrosamines), 히드록삼산(hydroxamic acids), 아조(azo) 염료, 옥심(oximes), 에폭사이드(epoxides), 방향족 니트로 화합물 및 1,2-벤즈이속사졸(1,2-benzisoxazole) 유도체와 같은 다양한 화합물을 환원시킴으로써 이들 약물에 대한 해독(Jordan 등 1999; Tatsumi 등 1983, 1986; Hirao 등 1994; Sugihara 등 1996)과 다중약물의 저항성을 극복하는데도 관여한다(McCrystal 등 1999). 한편 AO는 체내에서 내외인성 기질들의 산화 과정 중에 슈퍼옥사이드(superoxide)와 같은 ROS를 생성(Mira 등 1995; Kundu 등 2007)하는 것으로 알려져 있으며, 알코올성 간 손상을 유도(Shaw와 Jayatilleke, 1990)할 뿐만 아니라 고지방 식이에 의한 지방간의 생성에도 관여한다는 보고 (Conklin 등 2007; Weigert 등 2008)가 있다. 이러한 사실들을 고려해 볼 때 ROS와 관련된 질병의 발생에도 XO와 더불어 상당한 영향을 미칠 것으로 생각된다. 최근 AO의 mRNA는 내장 지방 조직(visceral adipose tissue)에서 피하 지방 조직(subcutaneous adipose tissue)에 비해 높게 발현되며(Vohl 등 2004; Weigert 등 2008), 비타민 A(vitamin A)의 알데하이드(aldehyde)로부터 지방세포(adipocyte)의 분화(Huang과 Ichikawa 1994; Mercader 등 2006)뿐만 아니라 간의 섬유화를 억제 (Neumeier 등 2006)하는 레티노산(retinoic acid)를 생성하는 것으로 알려져 있어, 비만의 조절과 간 손상의 예방 및 항암제 개발에 대한 표적효소 (Pryde 등 2010; Garattini와 Terao 2011)로 관심이 집중되고 있다. 즉, AO는 체내 독성물질의 해독에 관여하나 환자들이 복용중인 다양한 약물에 대하여는 그 효과를 감소시킬 수 있으며 특히, 음주로 인한 간 손상과 복부 비만을 유도하는데 관여할 뿐만 아니라 ROS 생성을 촉진하는데 관여함을 시사한다.Gout is a type of metabolic disease that causes arthritis. It is a disease caused by accumulation of uric acid crystals in the joint area due to hyperacidemia caused by excessive production or excretion of uric acid Chena et al. 2011). Uric acid is also the final metabolite of purine and is produced from xanthine by xanthine oxidase (XO). XO plays a major role in pathological conditions such as stress in the body (Oei et al., 1982; Urano et al., 1991; 90% of hyperactive acidosis is caused by insufficient excretion of kidneys and about 10% is caused by excessive production of uric acid (Wortmann 2002). Inadequate excretion of uric acid occurs in the urinary tract of the kidneys, while excessive uric acid is produced by increased purine or XO activity (Anzai et al. 2005; Caulfield et al. 2008). Recently, many antiepileptic drugs or XO inhibitors have been developed, but side effects such as digestive disorders, bone marrow suppression, kidney disorders, and hypersensitive syndromes are known (Horiuchi et al. 2000; Zhu et al. 2004; Robert and Terkeltaub 2003). It is similar in molecular weight to AO (aldehyde oxidase) XO and has complementary molecular chains such as flavin adenine dinucleotide (FAD), molybdenum and iron-sulfur and is closely related to in vivo action The specificity of the substrate has been reported to vary (Beedham 1987). AO is an enzyme that exists in large amounts in the cytoplasm of the liver, lungs, and kidneys (Critchley et al., 1992; Al-Salmy 2001) and contains aldehyde and nitrogen-containing heteroatoms including drugs such as methotrexate and antibiotics (famciclovir) (Clarke et al., 1995; Jordan et al., 1999; Kitamura et al., 1999). In the presence of electron donors, sulfoxides, N-oxides, nitrosamines ), Hydroxamic acids, azo dyes, oximes, epoxides, aromatic nitro compounds, and 1,2-benzisoxazole derivatives. (Jordan et al., 1999; Tatsumi et al., 1983; Hirao et al., 1994; Sugihara et al., 1996) and overcome the resistance of multiple drugs (McCrystal et al., 1999). AO is known to produce superoxide-like ROS during the oxidation of internal and external tough substrates in the body (Mira et al. 1995; Kundu et al. 2007) and induces alcoholic liver damage (Shaw and Jayatilleke, 1990) (Conklin et al., 2007; Weigert et al., 2008) have been reported to be involved in the production of fatty liver by high-fat diets. Considering these facts, it is thought that the occurrence of ROS-related diseases will have a considerable effect with XO. Recently, AO mRNA has been shown to be higher in visceral adipose tissue than in subcutaneous adipose tissue (Vohl et al., 2004; Weigert et al., 2008), the aldehyde of vitamin A (vitamin A) It is known to produce retinoic acid which inhibits the fibrosis of the liver as well as the differentiation of adipocytes (Huang and Ichikawa 1994; Mercader et al. 2006) Target enzymes for the development of anticancer drugs (Pryde et al. 2010; Garattini and Terao 2011) are attracting attention. In other words, AO is involved in the detoxification of the body toxins, but it can reduce the effects of various drugs being taken by the patients. In particular, it participates in inducing liver damage caused by alcohol and abdominal obesity, .
청미래덩굴(Smilax china L.)은 우리나라 산야에 널리 분포하는 활엽덩굴성 관목으로서, 이의 뿌리인 토복령은 트리테르페노이드 사포닌(triterpenoid saponins), 플리본(flavones), 스틸벤(stilbenes) 및 유기산(organic acids) 등이 주성분으로(Cheng과 Hua 2006) 한의학에서는 통풍치료(Song 등 2006; Shu 등 2006; Chen 등 2008; Wu 등 2010)에 쓰이며, 그 외에도 해독(Cheng과 Hua 2006), 항염, 항암 및 항산화 (Shu 등 2006; Li 등 2007) 효과가 있는 것으로 알려져 있다. 그러나 청미래덩굴의 잎은 예로부터 망개떡 제조에 사용해 왔으나 이에 대한 연구는 항균 및 항산화능 등 단편적인 연구(Choi 2004; Song 등 2006)가 있을 뿐이며 토복령이 가지는 통풍 예방효과 등 다양한 의약적 기능성에 대한 연구는 보이지 않는다. 예를 들어, 최근에 청미래덩굴 잎 추출물이 강한 항균력과 항산화력을 나타낸다는 보고가 있었고, 그 지표물질로 캠페롤-7-O-α-L-람노피라노사이드( kaempferol-7-O-α-L-rhamnopyranoside)와 캠페롤-3,7-O-α-L-다이람노피라노사이드(kaempferol-3,7-O-α-L-dirhamnopyranoside)가 분리된 바 있다(Cha & Lee 2007). 또한, 청미래덩굴 잎의 의약적 기능성과 관련하여 대한민국 등록특허공보 제10-1156636호에 청미래덩굴 잎 추출물을 유효성분으로 포함하는 혈관질환의 예방 또는 치료용 약학 조성물이 개시되어 있을 뿐이다. 청미래덩굴의 잎과 줄기 및 뿌리에는 폴리페놀(polyphenol) 화합물 중에서 특히 플라보노이드(flavonoid) 성분과 사포닌(saponin) 성분들이 풍부하게 함유되어 있으며 이들 성분들이 상기에서 언급한 바와 같은 청미래 덩굴이 가지고 있는 다양한 기능성과 최근에 발표되고 있는 항바이러스, 항비만, 청혈, 뇌신경보호 작용 등에도 관여하는 것으로 알려져 있다. 청미래덩굴의 잎, 줄기 및 뿌리에 함유되어 있는 플라보노이드(flavonoid) 류와 사포닌(saponin) 류는(Ruan 등 2002; Xu 등 2008) 대부분이 당류와 결합한 배당체(glycoside)의 형로 존재하나, 유리의 아글리폰(aglycone) 형태의 것이 우수한 기능성을 가지는 것으로 보고되어 있으며 이는 결합한 당류 부분이 반응에 장애를 주는 때문으로 알려져 있다(Hamzeh-Mivehroud 2013). 또한, 퀘라세틴(qurecetin), 루틴(rutin)과 같은 플라보노이드는 XO의 활성을 억제함으로써 통풍을 예방하는 효과가 알려져 있다(Zhu 등 2004). Smilax china L.) is a broad-leaved shrub that is widely distributed in mountainous areas of Korea. Its roots, toupengyong, contain triterpenoid saponins, flavones, stilbenes and organic acids. (Cheng and Hua 2006). In Oriental medicine, gout treatment 2006; Shu et al. 2006; Chen et al. 2008; Wu et al. 2010), and it is known that it has effects of detoxification (Cheng and Hua 2006), anti-inflammation, anti-cancer and antioxidant (Shu et al. 2006; Li et al. 2007). However, the leaves of Cheongmaea vine have been used for the manufacture of manggeok for some time. 2006), and studies on various medicinal functionalities such as the prevention of gout in Tohoku Bokryeong are not found. For example, it has recently been reported that the extracts of Cheongpyeongrae vine leaf have strong antimicrobial activity and antioxidant ability, and camphorol-7-O-α-L-rhamopyranoside (kaempferol-7-O-α L-rhamnopyranoside and camerol-3,7-O- alpha -L-kaempferol-3,7-O-alpha-L-dirhamnopyranoside have been isolated (Cha & Lee 2007) . In addition, in relation to the medicinal function of Cheongmyeongvin leaf, Korean Patent Registration No. 10-1156636 discloses a pharmaceutical composition for preventing or treating vascular diseases comprising Cheongmyeongvin leaf extract as an active ingredient. The leaves, stems and roots of Cheongmaea vine are abundant in polyphenol compounds, especially flavonoid and saponin components, and these components have various functionalities such as those mentioned above And the recently announced antiviral, anti-obesity, encephalopathy, cranial nerve protection is also known to be involved. Flavonoids and saponins contained in the leaves, stems and roots of Cheongmaea vine (Ruan et al. 2002; Xu 2008). It is reported that most of them are in the form of glycosides combined with saccharides, but free aglycone forms have been reported to have excellent functionality, which is believed to be due to the disruption of the bound saccharide moiety (Hamzeh-Mivehroud 2013). In addition, flavonoids such as qurecetin and rutin have been known to prevent gout by inhibiting the activity of XO (Zhu et al. 2004).
그러나, 청미래덩굴 잎은 특유의 신맛 등 관능적 특성이 떨어져 차와 같은 식품으로 대중화되기에는 한계가 있다. 또한, 청미래덩굴 잎의 추출물은 청미래덩굴 뿌리 추출물에 비해 일반적으로 생리활성이 매우 낮아 이를 기능성 식품 용도 내지 의약적 용도로 이용하기에는 한계가 있다.However, the Cheongmaea vine leaf has a unique sensory characteristic such as sour taste, which is limited to be popularized as a tea. In addition, the extract of Cheongmaea vine leaf is generally lower in physiological activity than that of Cheongmaea vine root extract, so that it is limited to use for functional foods or medicinal purposes.
본 발명은 이러한 배경하에 도출된 것으로서, 본 발명의 일 목적은 기능성 식품 내지 의약품 용도로 사용될 수 있도록 생리활성이 향상된 발효 청미래덩굴 잎, 이의 제조방법 및 이의 용도를 제공하는데에 있다.Accordingly, it is an object of the present invention to provide a fermented blue-green leaf blade having improved physiological activity so that it can be used for functional foods or pharmaceuticals, a method for producing the fermented blue-green leaf, and a use thereof.
또한, 본 발명의 다른 목적은 기능성 식품 내지 의약품 용도로 사용될 수 있는 발효 청미래덩굴 잎의 추출물, 이의 제조방법 및 이의 구체적인 용도를 제공하는데에 있다. It is another object of the present invention to provide an extract of Fermented Blueberry, which can be used for functional foods or pharmaceuticals, a process for producing the extract, and a specific use thereof.
본 발명의 발명자는 청미래덩굴 잎을 다양한 미생물로 발효시켜 발효 청미래덩굴 잎을 제조하고, 특정 미생물로 발효시킨 청미래덩굴 잎에서 관능적 특성이 개선되고, 이의 추출물이 향상된 생리활성을 가진다는 점을 발견하고 본 발명을 완성하였다.The inventors of the present invention have found that fermented Cheongmyeonghwinae leaves are fermented by various microorganisms to produce Cheongmaea vine leaves, and the sensory characteristics are improved in the Cheongmyeongvle leaf which is fermented with a specific microorganism, and its extract has improved physiological activity Thus completing the present invention.
본 발명의 일 목적을 해결하기 위하여, 본 발명은 아스퍼질러스속 미생물로 청미래덩굴 잎을 발효시켜 얻은 산물로서, 상기 발효에 의해 발효 전보다 향상된 잔틴 옥시다아제(Xanthine oxidase) 억제 활성 또는 알데하이드 옥시다아제(aldehyde oxidase) 억제 활성을 나타내는 것을 특징으로 하는 발효 청미래덩굴 잎을 제공한다. 또한, 본 발명은 (a) 청미래덩굴 잎에 아스퍼질러스속 미생물 배양액을 접종하고 청미래덩굴 잎을 발효시키는 단계; 및 (b) 발효된 청미래덩굴 잎을 건조하는 단계를 포함하고, 상기 발효된 청미래덩굴 잎은 발효 전보다 향상된 잔틴 옥시다아제(Xanthine oxidase) 억제 활성 또는 알데하이드 옥시다아제(aldehyde oxidase) 억제 활성을 나타내는 것을 특징으로 하는 발효 청미래덩굴 잎의 제조방법을 제공한다. 또한, 본 발명은 발효 청미래덩굴 잎의 용도로서 아스퍼질러스속 미생물에 의해 발효된 청미래덩굴 잎을 포함하는 발효 차를 제공한다. 또한, 본 발명은 발효 청미래덩굴 잎의 다른 용도로서 아스퍼질러스속 미생물에 의해 발효된 청미래덩굴 잎의 물 추출액을 포함하는 기능성 음료를 제공한다.
In order to solve one object of the present invention, the present invention provides a product obtained by fermenting a Cynomolgus japonica leaf with an Aspergillus microorganism, wherein the fermentation enhances the activity of inhibiting xanthine oxidase or aldehyde oxidase, Inhibiting activity of the fermented Cynthia raspberry leaf. (A) inoculating an Aspergillus microflora culture solution to the leaves of Cheongpyongjae, and fermenting the Cheongmyeonghwa leaf; And (b) drying the fermented Cyanoma vine leaf, wherein the fermented Cyanoma vine leaf exhibits an activity of inhibiting xanthine oxidase or aldehyde oxidase, which is more improved than before fermentation The present invention also provides a method for producing fermented blue-green crab leaves. In addition, the present invention provides a fermented tea comprising a Cheongmaea vine leaf fermented by an aspergillus microorganism as a fermented Cheongmyeong vine leaf. In addition, the present invention provides a functional beverage comprising a water extract of Cheongmyeongvin leaf fermented by an aspergillus microorganism as another use of fermented Cheongmyeong vine leaf.
본 발명의 다른 목적을 해결하기 위하여, 본 발명은 아스퍼질러스속 미생물에 의해 발효된 청미래덩굴 잎으로부터 추출되고, 잔틴 옥시다아제(Xanthine oxidase) 억제 활성 또는 알데하이드 옥시다아제(aldehyde oxidase) 억제 활성을 나타내는 발효 청미래덩굴 잎 추출물을 제공한다. 또한, 본 발명은 (a) 청미래덩굴 잎에 아스퍼질러스속 미생물 배양액을 접종하고 청미래덩굴 잎을 발효시키는 단계; (b) 발효된 청미래덩굴 잎을 건조하는 단계; 및 (c) 상기 건조된 발효 청미래덩굴 잎으로부터 추출물을 추출하는 단계를 포함하는 발효 청미래덩굴 잎 추출물의 제조방법을 제공한다. 또한, 본 발명은 발효 청미래덩굴 잎 추출물의 용도로서 발효 청미래덩굴 잎 추출물 또는 이의 분획물을 포함하는 통풍, 요통, 과요산혈증 또는 과요산뇨증의 예방 또는 치료용 조성물을 제공한다. 또한, 본 발명은 발효 청미래덩굴 잎 추출물의 용도로서 발효 청미래덩굴 잎 추출물 또는 이의 분획물을 포함하는 간 손상 예방 또는 치료용 조성물을 제공한다. 또한, 본 발명은 발효 청미래덩굴 잎 추출물의 용도로서 발효 청미래덩굴 잎 추출물 또는 이의 분획물을 포함하는 중성지방 또는 콜레스테롤 저하용 조성물을 제공한다. 이때, 상기 조성물은 약학 조성물 또는 식품 조성물에서 선택된 어느 하나인 것을 특징으로 한다.In order to solve the other object of the present invention, the present invention provides a fermented bluefin tuna extract, which is extracted from a bluefin tuna vine leaf fermented by an aspergillus microorganism and exhibits an activity of inhibiting xanthine oxidase or aldehyde oxidase, Leaf extract is provided. (A) inoculating an Aspergillus microflora culture solution to the leaves of Cheongpyongjae, and fermenting the Cheongmyeonghwa leaf; (b) drying the fermented blue-green crab leaf; And (c) extracting the extract from the dried fermented Cynthia sinensis leaf. The present invention also provides a composition for prevention or treatment of gout, low back pain, hyperlipidemia or hyperuricemia, comprising fermented Cheong pyung vine leaf extract or fractions thereof as an application of fermented Cheongmyeongvin leaf extract. The present invention also provides a composition for prevention or treatment of liver injury comprising fermented Cynomolgus thunbergii leaf extract or a fraction thereof, as an application of fermented Cynomolgus sinensis leaf extract. Further, the present invention provides a composition for neutral fat or cholesterol lowering, comprising fermented Cynomolgus niger leaf extract or a fraction thereof, as an application of fermented Cyanophyceae leaf extract. Here, the composition may be any one selected from a pharmaceutical composition or a food composition.
본 발명에 따른 발효 청미래덩굴 잎은 아스퍼질러스속 미생물로 발효되어 발효 전보다 향상된 잔틴 옥시다아제(Xanthine oxidase) 억제 활성 또는 알데하이드 옥시다아제(aldehyde oxidase) 억제 활성을 나타낸다. 또한, 본 발명에 따른 발효 청미래덩굴 잎은 발효 전보다 향상된 관능적 특성을 나타낸다. 따라서, 본 발명에 따른 발효 청미래덩굴 잎으로 제조한 발효 차나 기능성 음료는 상업적 품질이 우수하고 시음자의 건강을 유지시키거나 개선하는데에 유리하다. 또한, 본 발명에 따른 발효 청미래덩굴 잎의 추출물 또는 이의 분획물은 통풍, 간 손상, 비만, 고지혈증, 지방간 등을 예방, 개선 또는 치료하기 위한 의약품 소재 또는 기능성 식품 소재로 매우 유용하다.According to the present invention, the fermented Cynthia latifolia leaf is fermented into an aspergillus microorganism and exhibits an activity of inhibiting xanthine oxidase or aldehyde oxidase, which is more improved than before fermentation. In addition, the fermented blueberry leaf of the present invention exhibits improved sensory properties before fermentation. Therefore, the fermented tea or the functional beverage produced from the fermented green tea leaf of the present invention has an excellent commercial quality and is advantageous for maintaining or improving the health of the tasting person. In addition, the fermented green tea leaf extract or its fraction according to the present invention is very useful as a pharmaceutical material or a functional food material for preventing, ameliorating or treating gout, liver damage, obesity, hyperlipemia, fatty liver and the like.
도 1은 Aspergillus oryzae KFRI 995로 청미래덩굴 잎을 발효하여 얻은 발효 청미래덩굴 잎의 발효 시간에 따른 총 폴리페놀(TP) 함량 및 총 플라보노이드(TF) 함량 측정 결과를 나타낸 그래프이다.도 1은 Aspergillus oryzae KFRI 995로 청미래덩굴 잎을 발효하여 얻은 발효 청미래덩굴 잎의 발효 시간에 따른 총 폴리페놀(TP) 함량 및 총 플라보노이드(TF) 함량 측정 결과를 나타낸 그래프이다. 데이터는 3회 측정 평균값과 표준편차로 나타내었으며, 서로 다른 문자(A~D, a~d)는 5% 수준에서 유의성이 있음을 나타낸다.
도 2는 Aspergillus oryzae KFRI 995로 청미래덩굴 잎을 발효하여 얻은 발효 청미래덩굴 잎의 발효 시간에 따른 전자공여능(Electron donating ability, EDA) 측정 결과를 나타낸 그래프이고, 도 3은 Aspergillus oryzae KFRI 995로 청미래덩굴 잎을 발효하여 얻은 발효 청미래덩굴 잎의 발효 시간에 따른 철 환원력(Ferric reducing antioxidant power, FRAP) 측정 결과를 나타낸 그래프이다. 데이터는 3회 측정 평균값과 표준편차로 나타내었으며, 서로 다른 문자(A~C)는 5% 수준에서 유의성이 있음을 나타낸다.
도 4는 Aspergillus oryzae KFRI 995로 청미래덩굴 잎을 발효하여 얻은 발효 청미래덩굴 잎의 발효 시간에 따른 지질과산화 억제활성(Lipid peroxidation inhibitory activity, LPOIA) 측정 결과를 나타낸 그래프이다. 데이터는 3회 측정 평균값과 표준편차로 나타내었으며, 서로 다른 문자(A~B)는 5% 수준에서 유의성이 있음을 나타낸다.
도 5는 발효 청미래덩굴 잎 및 비발효 청미래덩굴 잎을 에탄올로 추출하고 에탄올 추출물을 에틸아세테이트로 분획하였을 때의 추출 수율을 나타낸 그래프이다.
도 6은 발효 청미래덩굴 잎 및 비발효 청미래덩굴 잎의 에탄올 추출물 및 이의 에틸아세테이트 가용성 분획물에 존재하는 총 폴리페놀 함량을 나타낸 그래프이고, 도 7은 발효 청미래덩굴 잎 및 비발효 청미래덩굴 잎의 에틸아세테이트 가용성 분획물에 존재하는 총 플라보노이드 함량 : 총 폴리페놀 함량의 비를 백분율로 나타낸 것이다.Figure 1 shows the Aspergillus (TP) content and total flavonoid (TF) content of fermented Cynomolgus sinensis leaf obtained by fermentation of Cheongmyeongvin leaf with oryzae KFRI 995. Figure 1 is a graph showing the results of measurement of total polyphenol (TP) content and TF content with fermentation time of Aspergillus (TP) content and total flavonoid (TF) content of Fermented Cynomolgus sinensis leaf obtained by fermentation of Cheongmyeongvle leaf with oryzae KFRI 995 according to fermentation time. Data are expressed as mean and standard deviation of three measurements, and different letters (A to D, a to d) indicate significance at the 5% level.
FIG. 2 shows the Aspergillus FIG. 3 is a graph showing the results of measuring the electron donating ability (EDA) according to the fermentation time of the fermented blueberry leaf vine leaves obtained by fermenting Cheongmyeongvin leaf with oryzae KFRI 995, FIG. 3 is a graph showing the results of Aspergillus (ferric reducing antioxidant power, FRAP) according to the fermentation time of the fermented Cynomolgus sinensis leaf obtained by fermenting Cheongmyeongvin leaf with oryzae KFRI 995. Data are presented as mean and standard deviation of three measurements, and different letters (A to C) indicate significance at the 5% level.
Figure 4 shows the Aspergillus (Lipid peroxidation inhibitory activity (LPOIA)) according to the fermentation time of fermented Cynomolgus sinensis leaf obtained by fermenting Cynomolgus sinensis leaf with oryzae KFRI 995. Data are presented as mean and standard deviation of three measurements, and different letters (A to B) indicate significance at the 5% level.
FIG. 5 is a graph showing the extraction yields of ethanol-extracted and ethanol-extracted fractions of fermented Cyanophyta and non-fermented Cyanoma vine leaves.
FIG. 6 is a graph showing the total polyphenol contents in the ethanol extracts of fermented Cheongpyong vine leaves and non-fermented Cheongmyeon vine leaves and their ethyl acetate soluble fractions, and FIG. 7 is a graph showing the total polyphenol contents in fermented Cheongpyong vine leaves and ethyl acetate The percentage of total flavonoid content: total polyphenol content present in the soluble fraction is expressed as a percentage.
이하, 본 발명에서 사용한 용어를 설명한다.Hereinafter, terms used in the present invention will be described.
본 발명에서 사용되는 용어 "예방"은 본 발명의 조성물의 투여로 특정 질환의 증상을 억제시키거나 진행을 지연시키는 모든 행위를 의미한다.As used herein, the term "prophylactic " means any act that inhibits the symptoms of a particular disease or delays the progress of the disease upon administration of the composition of the present invention.
본 발명에서 사용되는 용어 "치료"는 본 발명의 조성물의 투여로 특정 질환의 증상을 호전 또는 이롭게 변경시키는 모든 개선 행위를 의미한다.The term "treatment" as used herein refers to any improvement action that improves or alleviates the symptoms of a particular disease upon administration of the composition of the present invention.
본 발명에서 사용되는 용어 "투여"는 임의의 적절한 방법으로 개체에 소정의 본 발명의 조성물을 제공하는 것을 의미한다. 이때, 개체는 본 발명의 조성물을 투여하여 특정 질환의 증상이 호전될 수 있는 질환을 가진 인간, 원숭이, 개, 염소, 돼지 또는 쥐 등 모든 동물을 의미한다.
The term "administering" as used herein is meant to provide any desired composition of the invention to an individual by any suitable method. The term " individual " means any animal such as a human, a monkey, a dog, a goat, a pig, or a mouse having a disease in which symptoms of a specific disease can be improved by administering the composition of the present invention.
이하, 본 발명을 상세히 설명한다.
Hereinafter, the present invention will be described in detail.
본 발명의 일 측면은 유용한 생리활성을 나타내는 발효 청미래덩굴 잎, 이의 제조방법 및 이의 용도를 제공하는데에 있다.One aspect of the present invention is to provide a fermented blueberry leaf which exhibits useful physiological activity, a method for its production, and its use.
본 발명에 따른 발효 청미래덩굴 잎은 아스퍼질러스속 미생물로 청미래덩굴 잎을 발효시켜 얻은 산물로서 상기 발효에 의해 발효 전보다 향상된 잔틴 옥시다아제(Xanthine oxidase) 억제 활성 또는 알데하이드 옥시다아제(aldehyde oxidase) 억제 활성을 나타낸다. 이때, 아스퍼질러스속 미생물은 아스퍼질러스 카와치(Aspergillus kawachii), 아스퍼질러스 아와모리(Aspergillus awamori), 아스퍼질러스 오리재(Aspergillus oryzae), 아스퍼질러스 소재(Aspergillus sojae) 또는 아스퍼질러스 우사미(Aspergillus usamii)에서 선택된 하나 이상으로 구성될 수 있고, 청미래덩굴 잎의 생리활성을 극대화하는 측면에서 아스퍼질러스 오리재(Aspergillus oryzae)인 것이 바람직하다. 또한, 상기 아스퍼질러스 오리재(Aspergillus oryzae)는 Aspergillus oryzae KACC 40247 균주, Aspergillus oryzae KFRI 995 균주, Aspergillus oryzae var. brunneus KACC 44823 균주 또는 Aspergillus oryzae var effuses KACC 44990 균주에서 선택되는 하나 이상으로 구성될 수 있으나, 이에 한정되는 것은 아니다. 예를 들어, 아스퍼질러스 오리재(Aspergillus oryzae)는 기탁번호가 KCCM 12042, KCCM 12089, KCCM 12698, KCCM 32345, KCCM 60166, KCCM 60241 등에서 선택되는 하나 이상으로도 구성될 수 있다. 또한, 아스퍼질러스속 미생물의 발효 기질로 이용되는 청미래덩굴 잎은 발효 효율 등을 고려할 때 분말 형태인 것이 바람직하고, 이때 상기 분말의 입도 크기는 입도 크기가 20~100 메쉬(mesh)인 것이 바람직하고 40~80 메쉬(mesh)인 것이 더 바람직하나, 이에 한정되는 것은 아니다. 또한, 발효 청미래덩굴 잎이 후술하는 발효 차로 이용되거나 추출 등에 의해 기능성 음료의 소재로 이용될 때 사용의 편의성 및 추출 효율 등을 고려하는 측면에서 아스퍼질러스속 미생물에 의해 발효된 청미래덩굴 잎도 분말 형태인 것이 바람직하다.
According to the present invention, the fermented Cyanophylla leaf is an aspergillus microorganism, which is a product obtained by fermenting a Cyanoma leaf blade, and exhibits an activity of inhibiting xanthine oxidase or aldehyde oxidase by fermentation. In this case, Aspergillus seusok microorganism Aspergillus Kawachi (Aspergillus kawachii , Aspergillus awamori , Aspergillus oryzae ), aspergillus ( Aspergillus that the sojae) or Aspergillus Usami (Aspergillus usamii) Aspergillus duck material (Aspergillus oryzae) in the side can be composed of one or more selected, to maximize the biological activity of the leaves from Smilax China it is preferred. In addition, the Aspergillus oryzae may be Aspergillus oryzae KACC 40247 strain, Aspergillus oryzae KFRI 995 strain, Aspergillus There are oryzae . brunneus KACC 44823 strain or Aspergillus oryzae var effuses KACC 44990 strain, but the present invention is not limited thereto. For example, Aspergillus oryzae may also be constituted by one or more selected from the deposit numbers KCCM 12042, KCCM 12089, KCCM 12698, KCCM 32345, KCCM 60166, KCCM 60241 and the like. In addition, it is preferable that the Cheongmaea vine leaf used as a fermentation substrate of the microorganism of the aspergillus is powdery in consideration of the fermentation efficiency and the like. The particle size of the powder is preferably 20 to 100 mesh, More preferably 40 to 80 mesh, but is not limited thereto. In addition, in view of convenience in use and extraction efficiency when the fermented blueberry leaf is utilized as a fermentation tea to be described later or used as a functional beverage by the extraction or the like, the Cheongmyeong vine leaf fermented by aspergillus microorganisms is also in the form of powder .
본 발명에 따른 발효 청미래덩굴 잎의 제조방법은 (a) 청미래덩굴 잎에 아스퍼질러스속 미생물 배양액을 접종하고 청미래덩굴 잎을 발효시키는 단계; 및 (b) 발효된 청미래덩굴 잎을 건조하는 단계를 포함한다. 이때, 상기 발효된 청미래덩굴 잎은 발효 전보다 향상된 잔틴 옥시다아제(Xanthine oxidase) 억제 활성 또는 알데하이드 옥시다아제(aldehyde oxidase) 억제 활성을 나타낸다. 본 발명에 따른 발효 청미래덩굴 잎의 제조방법에서 사용되는 아스퍼질러스속 미생물은 아스퍼질러스 카와치(Aspergillus kawachii), 아스퍼질러스 아와모리(Aspergillus awamori), 아스퍼질러스 오리재(Aspergillus oryzae), 아스퍼질러스 소재(Aspergillus sojae) 또는 아스퍼질러스 우사미(Aspergillus usamii)에서 선택된 하나 이상으로 구성될 수 있고, 청미래덩굴 잎의 생리활성을 극대화하는 측면에서 아스퍼질러스 오리재(Aspergillus oryzae)인 것이 바람직하다. 또한, 상기 아스퍼질러스 오리재(Aspergillus oryzae)는 Aspergillus oryzae KACC 40247 균주, Aspergillus oryzae KFRI 995 균주, Aspergillus oryzae var. brunneus KACC 44823 균주 또는 Aspergillus oryzae var effuses KACC 44990 균주에서 선택되는 하나 이상으로 구성될 수 있으나, 이에 한정되는 것은 아니다. 예를 들어, 아스퍼질러스 오리재(Aspergillus oryzae)는 기탁번호가 KCCM 12042, KCCM 12089, KCCM 12698, KCCM 32345, KCCM 60166, KCCM 60241 등에서 선택되는 하나 이상으로도 구성될 수 있다. 또한, 본 발명에 따른 발효 청미래덩굴 잎의 제조방법은 바람직하게는 상기 (a) 단계 이전에 청미래덩굴 잎을 건조 후 분쇄하는 단계를 더 포함할 수 있다. 예를 들어, 청미래덩굴 잎은 아스퍼질러스속 미생물에 의한 청미래덩굴 잎의 발효 효율 등을 고려할 때 바람직하게는 입도 크기가 20~100 메쉬(mesh)인 분말 형태로 분쇄될 수 있고, 더 바람직하게는 입도 크기가 40~80 메쉬(mesh)인 분말 형태로 분쇄될 수 있으나, 이에 한정되는 것은 아니다. 또한, 본 발명에 따른 발효 청미래덩굴 잎의 제조방법은 청미래덩굴 잎의 발효시 오염 등을 방지하기 위해 상기 (a) 단계 이전에 청미래덩굴 잎의 분말을 멸균하는 단계를 더 포함할 수 있다. 상기 멸균 방법은 공지의 다양한 방법에서 선택될 수 있으며, 예를 들어 청미래덩굴 잎의 분말에 소정의 물을 첨가하고 혼합한 후, 소정의 온도로 가열하는 것으로 구성될 수 있다. 이때, 물의 첨가량은 크게 제한되지 않으나, 청미래덩굴 잎의 분말과의 혼합 용이성 및 멸균의 효율성 등을 고려할 때 청미래덩굴 잎 분말 중량 대비 1~3배인 것이 바람직하고, 1.5~2.5배인 것이 더 바람직하다. 또한, 멸균 시 가열 온도는 100~125℃인 것이 바람직하고, 110~125℃인 것이 더 바람직하다. 또한, 멸균 시 가열 시간은 청미래덩굴 잎 분말의 상태 변화 및 멸균 효과를 담보하는 측면에서 10~120분인 것이 바람직하고, 30~90분인 것이 더 바람직하며, 40~80분인 것이 가장 바람직하다. 다만, 본 발명에 따른 청미래덩굴 잎의 발효가 고상 발효에 의해 이루어지는 경우 청미래덩굴 잎의 발효 이전에 청미래덩굴 잎을 멸균하는 단계를 생략할 수 있다.The method of manufacturing fermented Cheong pyramid according to the present invention comprises the steps of: (a) inoculating an Aspergillus sp. And (b) drying the fermented blue-green vine leaves. At this time, the fermented Rhododendron vine leaf shows an activity of inhibiting xanthine oxidase or aldehyde oxidase, which is higher than that before fermentation. Aspergillus seusok microorganism used in the manufacturing method of fermented Smilax China leaf according to the invention Aspergillus Kawachi (Aspergillus kawachii , Aspergillus awamori , Aspergillus oryzae ), aspergillus ( Aspergillus sojae) or Aspergillus Usami (Aspergillus usamii) can be composed of one or more selected from, Aspergillus duck material (Aspergillus in terms of maximizing the biological activities of Smilax China leaf oryzae . In addition, the Aspergillus oryzae ) is Aspergillus oryzae KACC 40247 strain, Aspergillus oryzae KFRI 995 strain, Aspergillus There are oryzae . brunneus KACC 44823 strain or Aspergillus oryzae var effuses KACC 44990 strain, but the present invention is not limited thereto. For example, Aspergillus oryzae may also be constituted by one or more selected from the deposit numbers KCCM 12042, KCCM 12089, KCCM 12698, KCCM 32345, KCCM 60166, KCCM 60241 and the like. In addition, the method for producing the fermented blueberry leaf blade according to the present invention may further comprise a step of drying and crushing the Cheongmyeongvle leaf before the step (a). For example, when considering the efficiency of fermentation of Cheongmyeongvin leaf by aspergillus microorganisms, Cheongmaejinae can be pulverized into a powder having a particle size of 20 to 100 mesh, and more preferably, And may be pulverized into a powder having a particle size of 40 to 80 mesh, but is not limited thereto. In addition, the method of manufacturing the fermented Cheongmyeong vine leaf according to the present invention may further include a step of sterilizing the powder of Cheongmyeong vine leaf before the step (a) in order to prevent contamination during fermentation of the Cheongmyeongvle leaf. The sterilization method may be selected from a variety of known methods. For example, the sterilization may be performed by adding predetermined water to powders of Rhododendron vine leaf, mixing and heating the mixture to a predetermined temperature. In this case, the amount of water to be added is not limited, but it is preferably 1 to 3 times, more preferably 1.5 to 2.5 times, based on the weight of the powder of Crassulaceae, in consideration of easiness of mixing with the powder of Crassulaceae, and efficiency of sterilization. The sterilization temperature is preferably 100 to 125 ° C, more preferably 110 to 125 ° C. The heating time at the time of sterilization is preferably 10 to 120 minutes, more preferably 30 to 90 minutes, and most preferably 40 to 80 minutes in view of ensuring the change of state and sterilization effect of the green tea leaf powder. However, when the fermentation of Cheongmaea vine leaf according to the present invention is performed by solid phase fermentation, the step of sterilization of Cheongmaea vine leaf may be omitted before fermentation of Cheongmaea vine leaf.
본 발명에 따른 발효 청미래덩굴 잎의 제조방법에서 청미래덩굴 잎에 접종되는 아스퍼질러스속 미생물 배양액은 아스퍼질러스속 미생물을 액체 배지에 배양한 것으로서, 통상적으로 사용되는 종균액에 해당하는 개념이다. 이때, 상기 아스퍼질러스속 미생물 배양액의 미생물 농도는 적정 수준으로 조정되는 것이 바람직한데, 예를 들어 106 cells/㎖ 내지 1010 cells/㎖로 조정될 수 있고, 더 바람직하게는 107 cells/㎖ 내지 109 cells/㎖로 조정될 수 있다. 또한, 본 발명에 따른 발효 청미래덩굴 잎의 제조방법에서 청미래덩굴 잎에 접종되는 아스퍼질러스속 미생물 배양액의 접종량은 크게 제한되지 않으나 발효 공정의 경제성 및 청미래덩굴 잎의 생리활성을 극대화하는 측면을 고려할 때 청미래덩굴 잎 건조 중량 대비 1~15 부피%인 것이 바람직하고, 2~12 부피%인 것이 더 바람직하며, 4~10 부피%인 것이 가장 바람직하다. 또한, 청미래덩굴 잎을 아스퍼질러스속 미생물로 발효시키는 단계에서 청미래덩굴 잎은 총 수분 함량이 50~75%인 것이 바람직하고 60~75 중량%인 것이 더 바람직하며, 65~70%인 것이 가장 바람직하다. 청미래덩굴 잎의 총 수분 함량이 75%를 초과하는 경우 액상 발효에 가까워 발효 과정 중에 오염될 염려가 있고, 청미래덩굴 잎의 총 수분 함량이 50% 미만인 경우 아스퍼질러스속 미생물에 의한 발효가 원활하지 않을 수 있다. 청미래덩굴 잎의 총 수분 함량은 청미래덩굴 잎에 접종되는 아스퍼질러스속 미생물 배양액과 추가로 첨가되는 물 등에 의해 조정될 수 있다. 또한, 본 발명에 따른 발효 청미래덩굴 잎의 제조방법에서 청미래덩굴 잎의 발효는 발효 공정의 경제성 등 및 오염의 최소화 등을 고려할 때 고상 발효에 의해 이루어지는 것이 바람직하다. 또한, 본 발명에 따른 발효 청미래덩굴 잎의 제조방법에서 발효 온도는 사용되는 아스퍼질러스속 미생물의 종류 등에 따라 다양한 범위를 가질 수 있으며, 아스퍼질러스속 미생물의 최적 배양 온도 등을 고려할 때 20~35℃인 것이 바람직하고, 25~35℃인 것이 더 바람직하며, 25~32℃인 것이 가장 바람직하다. 또한, 본 발명에 따른 발효 청미래덩굴 잎의 제조방법에서 발효 시간은 청미래덩굴 잎의 발효 시 사용되는 아스퍼질러스속 미생물의 종류, 아스퍼질러스속 미생물 배양액의 접종량, 발효 온도 등에 따라 다양한 범위를 가질 수 있으며, 발효 청미래덩굴 잎의 관능적 특성, 발효 공정의 경제성 또는 청미래덩굴 잎의 생리활성을 극대화하는 측면을 고려할 때 5~30일인 것이 바람직하고, 5~20일 것이 더 바람직하며, 7~15일인 것이 가장 바람직하다.In accordance with the present invention, the culture medium of the aspergillus microorganism inoculated on the leaf of Cyperma javanica is the culture of the aspergillus microorganism in a liquid medium, which is a concept corresponding to a commonly used seed medium. At this time, it is preferable that the microorganism concentration of the aspherical microorganism culture liquid is adjusted to an appropriate level, for example, it may be adjusted to 10 6 cells / ml to 10 10 cells / ml, more preferably 10 7 cells / 10 < 9 > cells / ml. In addition, although the inoculation amount of the culture medium of the aspergillus microorganism inoculated on the Cheongmyeongvle leaf is not limited in the method of manufacturing the fermented Cheonggyejina vine leaf according to the present invention, considering the economical efficiency of the fermentation process and maximizing the physiological activity of the Cheongmyeongvle leaf It is preferably 1 to 15% by volume, more preferably 2 to 12% by volume, and most preferably 4 to 10% by volume based on the dry weight of the Cheongmyeongvin leaf. In addition, in the step of fermenting the Cheongmaia crane leaf as an aspherical microorganism, the total moisture content of the Cheongmaia crane leaf is preferably 50 to 75%, more preferably 60 to 75%, most preferably 65 to 70% Do. When the total moisture content of the Cheongmyeongvin leaf exceeds 75%, there is a risk of contamination during the fermentation process due to the near-liquid fermentation, and when the total moisture content of the Cheongmyeongvine leaf is less than 50%, the fermentation by the aspergillus microorganism is not smooth . The total moisture content of the Cheongmyeong vine leaves can be adjusted by the culture of the aspergillus microorganism inoculated on the Cheongmyeongvle leaf and the water added additionally. In addition, in the method of manufacturing fermented Cheong pyramid according to the present invention, it is preferable that the fermentation of Cheongmaea vine leaves is performed by solid phase fermentation considering economical efficiency of the fermentation process and minimization of contamination. The fermentation temperature may be varied depending on the kind of the aspherical microorganism to be used. In consideration of the optimal culture temperature of the aspergillus microorganism, the fermentation temperature may be 20 to 35 ° C More preferably 25 to 35 占 폚, and most preferably 25 to 32 占 폚. In addition, the fermentation time of the fermented blueberry leaf blade according to the present invention may be varied depending on the kind of aspergillus microorganism used for fermentation of Cheongmaea vine leaf, inoculation amount of aspergillus microorganism culture solution, fermentation temperature, and the like It is preferably 5 to 30 days, more preferably 5 to 20 days, most preferably 7 to 15 days, in view of maximizing the physiological activity of the fermentation process or the economical efficiency of the fermentation process or the sensory characteristics of the fermented green perilla leaf, desirable.
본 발명에 따른 발효 청미래덩굴 잎의 제조방법에서 아스퍼질러스속 미생물에 의해 발효된 청미래덩굴 잎은 열풍 건조 등 다양한 방법에 의해 건조되어 건조된 형태로 제공된다. 이때, 본 발명에 따른 발효 청미래덩굴 잎의 제조방법은 바람직하게는 발효된 청미래덩굴 잎을 건조하기 전에 멸균하는 단계를 더 포함할 수 있다. 발효된 청미래덩굴 잎을 건조하기 전에 멸균함으로써 추후의 오염을 방지할 수 있고 제품의 안전성을 도모할 수 있다. 상기 멸균 방법은 공지의 다양한 방법에서 선택될 수 있으며, 예를 들어 청미래덩굴 잎을 아스퍼질러스속 미생물로 발효시켜 얻은 산물을 오토클레이브 등을 이용하여 소정의 압력 및 소정의 온도로 가열하는 것으로 구성될 수 있다. 이때, 멸균 시 가열 온도는 100~125℃인 것이 바람직하고, 110~125℃인 것이 더 바람직하다. 또한, 멸균 시 가열 시간은 멸균 효과를 담보하는 측면에서 10~120분인 것이 바람직하고, 30~90분인 것이 더 바람직하며, 40~80분인 것이 가장 바람직하다.
In the method of manufacturing fermented blue-green algae according to the present invention, Cheongmaea vine leaf fermented by an aspergillus microorganism is dried and dried by various methods such as hot air drying. At this time, the method of manufacturing fermented Cyanophyta blade according to the present invention may further include a step of sterilizing fermented Cyanoma vine leaf before drying. The fermented Cheongmyeonghwa leaf is sterilized before it is dried, so that it is possible to prevent further contamination and ensure the safety of the product. The sterilization method may be selected from a variety of known methods. For example, the sterilization method may include heating a product obtained by fermenting a Korean blue-green leaf with an aspherical microorganism to a predetermined pressure and a predetermined temperature using an autoclave or the like . At this time, the heating temperature at the time of sterilization is preferably 100 to 125 ° C, more preferably 110 to 125 ° C. The sterilization time is preferably 10 to 120 minutes, more preferably 30 to 90 minutes, and most preferably 40 to 80 minutes in terms of ensuring the sterilization effect.
본 발명은 전술한 발효 청미래덩굴 잎의 일 용도로 발효 청미래덩굴 잎을 포함하는 발효 차를 제공한다. 상기 발효 청미래덩굴 잎을 포함하는 발효 차는 비발효 청미래덩굴 잎에 비해 향상된 관능적 특성, 잔틴 옥시다아제(Xanthine oxidase) 억제 활성 또는 알데하이드 옥시다아제(aldehyde oxidase) 억제 활성을 나타낸다. 또한, 본 발명에 따른 발효 차는 발효 청미래덩굴 잎 외에 상업적 제품 수준의 관능적 특성을 담보하는 측면에서 옥수수 뿌리 분말, 옥수수 줄기 분말, 꾸지뽕 분말 또는 가시오가피 분말에서 선택되는 하나 이상을 더 포함할 수 있다. 또한, 본 발명에 따른 발효 차는 사용의 편의성 측면에서 티백 형태로 제공될 수 있다. 상기 티백에는 발효 청미래덩굴 잎이 수용될 수 있고, 선택적으로 옥수수 뿌리 분말, 옥수수 줄기 분말, 꾸지뽕 분말 또는 가시오가피 분말에서 선택되는 하나 이상이 더 수용될 수도 있다.The present invention provides a fermented tea comprising a fermented green tea leaf blade for the purpose of the aforementioned fermented green tea leaf. The fermented tea containing the fermented Cynthia raspberry exhibits improved sensory properties, xanthine oxidase inhibitory activity or aldehyde oxidase inhibitory activity as compared to nonfermented Cynthia rybiaceae. In addition, the fermented tea according to the present invention may further include at least one selected from corn root powder, corn stover powder, coconut powder, or goethiose powder from the viewpoint of securing the sensory characteristics of a commercial product in addition to the fermented Cynthia raspberry. Further, the fermented tea according to the present invention may be provided in the form of a tea bag in terms of ease of use. The tea bag may contain a fermented green tea leaf, and may optionally contain one or more selected from corn root powder, corn stalk powder, gherkin powder or ganoderma powder.
또한, 본 발명은 전술한 발효 청미래덩굴 잎의 다른 용도로 발효 청미래덩굴 잎의 물 추출액을 포함하는 기능성 음료를 제공한다. 이때, 발효 청미래덩굴의 추출 용매인 물은 추출 효율을 고려할 때 열수인 것이 바람직하다. 또한, 본 발명에 따른 기능성 음료는 발효 청미래덩굴 잎의 물 추출액 외에 상업적 제품 수준의 관능적 특성을 담보하는 측면에서 옥수수 뿌리 추출액, 옥수수 줄기 추출액, 흑삼 추출액, 사양벌꿀, 비타민 또는 가시오가피 추출액에서 선택되는 하나 이상을 더 포함할 수 있다. 본 발명에 따른 기능성 음료는 통풍, 간 손상, 비만, 고지혈증, 지방간 등을 예방 또는 개선하는 효과가 있으므로, 시음자의 건강을 유지시키거나 개선하는데에 유리하다.
In addition, the present invention provides a functional beverage comprising a water extract of fermented Cynthia raspberry leaf for other uses of the above fermented Cynthia raspberry. At this time, it is preferable that water, which is an extraction solvent of the fermented bluefin tuna, is hydrothermal in consideration of extraction efficiency. In addition, the functional beverage according to the present invention can be used in the form of a corn root extract, a corn stalk extract, a black ginseng extract, a specimen honey, a vitamin or an extract from a sea tangle, in view of securing the sensory characteristics of a commercial product, Or more. The functional beverage according to the present invention has an effect of preventing or ameliorating gout, liver damage, obesity, hyperlipemia, fatty liver and the like, and thus is advantageous for maintaining or improving the health of the tasting person.
본 발명의 다른 측면은 유용한 생리활성을 나타내는 발효 청미래덩굴 잎 추출물, 이의 제조방법 및 이의 용도를 제공하는데에 있다. 이하, 전술한 발효 청미래덩굴 잎 추출물, 이의 제조방법 및 이의 용도와 동일한 내용은 기재를 생략한다.Another aspect of the present invention is to provide a fermented Cynomolgus thunbergii leaf extract showing useful physiological activity, a method for its production, and its use. Hereinafter, the same contents as those of the above-described Fermented Cyanophyceae leaf extract, its preparation method and its uses are not described.
본 발명에 따른 발효 청미래덩굴 잎 추출물은 아스퍼질러스속 미생물에 의해 발효된 청미래덩굴 잎으로부터 추출된 것으로서, 잔틴 옥시다아제(Xanthine oxidase) 억제 활성 또는 알데하이드 옥시다아제(aldehyde oxidase) 억제 활성을 나타낸다. 이때, 발효 청미래덩굴 잎 추출물은 바람직하게는 비발효 청미래덩굴 잎 추출물에 비해 향상된 잔틴 옥시다아제(Xanthine oxidase) 억제 활성 또는 알데하이드 옥시다아제(aldehyde oxidase) 억제 활성을 나타낸다. 또한, 본 발명에 따른 발효 청미래덩굴 잎 추출물은 아스퍼질러스속 미생물에 의해 청미래덩굴 잎이 발효되는 과정에서 생성된 다양한 생리활성 물질을 포함하고, 이로 인해 다양한 기능성을 가진다.According to the present invention, the fermented Cynthia rumulus leaf extract is extracted from Cheongmyeongvle leaf fermented by an Aspergillus microorganism and exhibits a xanthine oxidase inhibitory activity or an aldehyde oxidase inhibitory activity. In this case, the fermented Cynomolgus sinensis leaf extract preferably exhibits an enhanced inhibitory activity of xanthine oxidase or an aldehyde oxidase inhibitory activity compared with non-fermented Cynthia sinensis extract. In addition, the fermented Cyanophyceae leaf extract according to the present invention contains various physiologically active substances produced during the fermentation of Cyanobacterium japonica by the aspergillus microorganisms, and thus has various functionalities.
본 발명에 따른 발효 청미래덩굴 잎 추출물의 제조방법은 (a) 청미래덩굴 잎에 아스퍼질러스속 미생물 배양액을 접종하고 청미래덩굴 잎을 발효시키는 단계; (b) 발효된 청미래덩굴 잎을 건조하는 단계; 및 (c) 상기 건조된 발효 청미래덩굴 잎으로부터 추출물을 추출하는 단계를 포함한다. 이 중 (a) 단계 및 (b) 단계는 발효 청미래덩굴 잎의 제조방법에서 서술한 내용과 동일하므로 설명을 생략한다. 본 발명에 따른 발효 청미래덩굴 잎 추출물은 당업계에 공지된 통상의 추출 방법, 예를 들어 용매 추출법을 사용하여 제조될 수 있다. 용매 추출법을 이용한 발효 청미래덩굴 잎 추출물 제조시 사용되는 추출 용매는 물, 탄소 수가 1 내지 4인 저급 알코올(예를 들면, 메탄올, 에탄올, 프로판올 및 부탄올) 또는 함수 저급 알코올, 프로필렌글리콜, 1,3-부틸렌글리콜, 글리세린, 아세톤, 디에틸에테르, 에틸아세테이트, 부틸아세테이트, 디클로로메탄, 클로로포름, 헥산 및 이들의 혼합물로 구성된 군으로부터 선택될 수 있고, 이중 물, 알코올 또는 이들의 혼합물에서 선택되는 것이 바람직하다. 추출 용매로 물을 사용하는 경우 물은 열수인 것이 바람직하다. 예를 들어 본 발명에 따른 발효 청미래덩굴 잎 추출물은 발효 청미래덩굴 잎에 발효 청미래덩굴 잎 중량 대비 5~200배 부피의 물을 가하고 80~120℃에서 5분 내지 1시간 동안 환류 추출 후 여과하고 동결건조하여 얻을 수 있다. 또한, 추출 용매로 알코올을 사용하는 경우 알코올은 탄소 수가 1 내지 4인 저급 알코올인 것이 바람직하고, 저급 알코올은 메탄올 또는 에탄올에서 선택되는 것이 더 바람직하다. 또한, 추출 용매로 함수 저급 알코올을 사용하는 경우 함수 저급 알코올은 에탄올 농도가 60~90%인 것이 바람직하다. 또한, 본 발명의 발효 청미래덩굴 잎 추출물은 발효 청미래덩굴 잎의 열수, 알코올 또는 함수 알코올 추출물에서 에틸아세테이트 가용성 성분만을 분획한 분획 추출물을 포함한다. 한편, 본 발명의 발효 청미래덩굴 잎 추출물은 상기한 추출 용매뿐만 아니라, 다른 추출 용매를 이용하여도 실질적으로 동일한 효과를 나타내는 추출물이 얻어질 수 있다는 것은 당업자에게 자명한 것이다.
The method of manufacturing a fermented Cheong pyramid vine leaf extract according to the present invention comprises the steps of: (a) inoculating an Aspergillus sp. (b) drying the fermented blue-green crab leaf; And (c) extracting the extract from the dried fermented Cynthia sinensis leaf. The steps (a) and (b) are the same as those described in the preparation method of fermented blue-green crab leaves, and the description thereof will be omitted. The fermented Cyanophyta leaf extract according to the present invention can be produced by a conventional extraction method known in the art, for example, a solvent extraction method. The extraction solvent used in preparing the fermented Cheongmaia vine leaf extract by the solvent extraction method is water, a lower alcohol having 1 to 4 carbon atoms (for example, methanol, ethanol, propanol and butanol) or a hydrated lower alcohol, propylene glycol, 1,3 -Butylene glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane and mixtures thereof, of which water, alcohol or mixtures thereof desirable. When water is used as the extraction solvent, the water is preferably hot water. For example, the fermented blueberry leaf vine leaf extract according to the present invention is prepared by adding 5 to 200 times as much water as fermented blueberry leaf vine leaves to the fermented blueberry leaf vine leaves, refluxing the extract at 80 to 120 ° C for 5 minutes to 1 hour, Dried. When an alcohol is used as an extraction solvent, the alcohol is preferably a lower alcohol having 1 to 4 carbon atoms, and the lower alcohol is more preferably selected from methanol or ethanol. In addition, when the functional lower alcohol is used as the extraction solvent, the ethanol concentration of the functional lower alcohol is preferably 60 to 90%. In addition, the fermented Cyanophyta crane leaf extract of the present invention includes fraction extracts obtained by fractionating only the ethyl acetate soluble component in the hydrothermal, alcohol, or hydroalcoholic extracts of fermented Cyanophyceae. It is apparent to those skilled in the art that the fermented Cyanophyta leaf extract of the present invention can be used not only in the above-mentioned extraction solvent, but also in other extracting solvents.
본 발명에 따른 발효 청미래덩굴 잎 추출물 또는 이의 분획물은 통풍, 요통, 과요산혈증 또는 과요산뇨증의 예방 또는 치료용 조성물의 유효성분으로 사용될 수 있다. 또한, 본 발명에 따른 발효 청미래덩굴 잎 추출물 또는 이의 분획물은 간 손상 예방 또는 치료용 조성물의 유효성분으로 사용될 수 있다. 이때, 상기 간 손상은 알코올 등 다양한 간 독성 물질에 유발되는 것으로서 간의 비대, 간의 기능 저하 등을 포함한다. 또한, 본 발명에 따른 발효 청미래덩굴 잎 추출물 또는 이의 분획물은 중성지방 또는 콜레스테롤 저하용 조성물의 유효성분으로 사용될 수 있다. 본 발명에 따른 발효 청미래덩굴 잎 추출물 또는 이의 분획물을 포함하는 중성지방 또는 콜레스테롤 저하용 조성물은 구체적으로 지방간, 고지질혈증, 고중성지방혈증, 고콜레스테롤혈증, 지질 관련 대사 증후군과 같은 지질 관련 대사성 질환, 비만 등을 예방 또는 치료하는데에 사용될 수 있다. 이때, 본 발명에 따른 상기 조성물 들은 사용 목적 내지 양상에 따라 약학 조성물 또는 식품 조성물 등으로 구체화될 수 있다.The fermented Cynomolgus sinensis leaf extract or its fractions according to the present invention can be used as an active ingredient of a composition for preventing or treating gout, back pain, hyperuricemia or hyperuricemia. In addition, the fermented Cynomolgus sinensis leaf extract or fractions thereof according to the present invention can be used as an active ingredient of a composition for preventing or treating liver injury. At this time, the liver damage is caused by various liver toxic substances such as alcohol and includes liver hypertrophy, liver liver function and the like. In addition, the fermented Cynomolgus sinensis leaf extract or its fractions according to the present invention can be used as an active ingredient of a composition for neutral fat or cholesterol lowering. The composition for neutral fat or cholesterol lowering, which comprises the fermented Celestine vine leaf extract or the fraction thereof, according to the present invention is a lipid-related metabolic disease such as fatty liver, hyperlipidemia, hypertriglyceridemia, hypercholesterolemia, lipid-related metabolic syndrome , Obesity, and the like. At this time, the compositions according to the present invention may be formulated into a pharmaceutical composition, a food composition or the like according to the intended use or aspect.
본 발명의 발효 청미래덩굴 잎 추출물(예를 들어 청미래덩굴 잎의 열수 추출물 또는 함수 알코올 추출물) 또는 이의 분획물(예를 들어 청미래덩굴 잎의 열수 추출물 또는 함수 알코올 추출물의 에틸아세테이트 가용성 분획물)을 유효성분으로 포함하는 약학 조성물에서 발효 청미래덩굴 잎 추출물 또는 이의 분획물의 함량은 크게 제한되지 않으며, 예를 들어 조성물 총 중량을 기준으로 0.1~99 중량%, 바람직하게는 0.5~50 중량%, 더 바람직하게는 1~30 중량%일 수 있다. 또한, 본 발명에 따른 약학 조성물은 발효 청미래덩굴 잎 추출물 또는 이의 분획물 외에 약학적으로 허용가능한 담체, 부형제 또는 희석제와 같은 첨가제를 더 포함할 수 있다. 발명의 약학 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. 또한, 본 발명의 약학적 조성물은 발효 청미래덩굴 잎 추출물 또는 이의 분획물 외에 항산화 활성, 콜레스테롤 저하 활성, 지질 저하 활성 또는 중성지방 저하 활성을 갖는 공지의 유효 성분을 1종 이상 더 함유할 수 있다. 본 발명의 약학 조성물은 통상의 방법에 의해 경구 투여를 위한 제형 또는 비경구 투여를 위한 제형으로 제제화될 수 있고, 제제화할 경우 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제될 수 있다. 경구 투여를 위한 고형 제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형 제제는 발효 청미래덩굴 잎 추출물 또는 이의 분획물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트(Calcium carbonate), 수크로스(Sucrose), 락토오스(Lactose) 또는 젤라틴 등을 섞어 조제될 수 있다. 또한, 단순한 부형제 이외에 마그네슘 스티레이트 탈크 같은 윤활제들도 사용될 수 있다. 경구 투여를 위한 액상 제제로는 현탁제, 내용액제, 유제 및 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조제제, 좌제가 포함될 수 있다. 비수성용제, 현탁용제로는 프로필렌글리콜(Propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다. 더 나아가 당 분야의 적정한 방법으로 또는 Remington's Pharmaceutical Science(최근판), Mack Publishing Company, Easton PA에 개시되어 있는 방법을 이용하여 각 질환에 따라 또는 성분에 따라 바람직하게 제제화할 수 있다. 본 발명의 약학 조성물은 목적하는 방법에 따라 인간을 포함한 포유류에 경구 투여되거나 비경구 투여될 수 있으며, 비경구 투여 방식으로는 피부 외용, 복강내주사, 직장내주사, 피하주사, 정맥주사, 근육내 주사 또는 흉부내 주사 주입방식 등이 있다. 본 발명의 약학 조성물의 투여량은 환자의 체중, 연령, 성별, 건강상태, 식이, 투여시간, 투여방법, 배설율 및 질환의 중증도에 따라 그 범위가 다양하다. 본 발명의 약학 조성물의 통상적인 1일 투여량은 크게 제한되지 않으나 바람직하게는 유효성분인 발효 청미래덩굴 잎 추출물 또는 이의 분획물을 기준으로 할 때 0.1 내지 1000 ㎎/㎏이고, 더 바람직하게는 1 내지 500 ㎎/㎏이며, 하루 1회 또는 수회로 나누어 투여될 수 있다.(For example, a hydrothermal extract or a hydroalcoholic extract of Cheongmahra vine leaf) or a fraction thereof (for example, a hydrothermal extract or a ethyl acetate-soluble fraction of a hydroalcoholic extract of Cheongpyeongvle leaf) of the present invention as an active ingredient The content of the fermented green tea leaf extract or fraction thereof in the pharmaceutical composition to be contained is not particularly limited and may be, for example, 0.1 to 99% by weight, preferably 0.5 to 50% by weight, more preferably 1 To 30% by weight. In addition, the pharmaceutical composition according to the present invention may further comprise additives such as a pharmaceutically acceptable carrier, excipient or diluent in addition to the fermented Celery root extract or fraction thereof. 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. In addition, the pharmaceutical composition of the present invention may contain at least one known active ingredient having antioxidative activity, cholesterol lowering activity, lipid lowering activity, or triglyceride lowering activity, in addition to the fermented Cyanophyceae leaf extract or its fractions. The pharmaceutical composition of the present invention can be formulated into a formulation for oral administration or parenteral administration by a conventional method, and can be formulated into a pharmaceutical composition such as a filler, an extender, a binder, a wetting agent, a disintegrant, Diluents or excipients. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient, for example starch, calcium carbonate ), Sucrose, lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate talc may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions and syrups. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. Examples of suppository bases include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like. Further, it can be suitably formulated according to each disease or ingredient, using appropriate methods in the art or by the method disclosed in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA. The pharmaceutical composition of the present invention may be administered orally or parenterally to a mammal including a human according to a desired method. Examples of the parenteral administration method include external dermal application, intraperitoneal injection, intramuscular injection, subcutaneous injection, intravenous injection, Intravenous injection or intra-thoracic injection. The dosage of the pharmaceutical composition of the present invention varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and disease severity. The typical daily dose of the pharmaceutical composition of the present invention is not particularly limited, but is preferably 0.1 to 1000 mg / kg, more preferably 1 to 1000 mg / kg, based on the active ingredient, 500 mg / kg, and can be administered once a day or divided into several times a day.
본 발명의 발효 청미래덩굴 잎 추출물 또는 이의 분획물을 유효 성분으로 포함하는 식품 조성물은 환제, 분말, 과립, 침제, 정제, 캡슐, 또는 액제 등의 형태를 포함하며, 구체적인 식품의 예로는 육류, 소시지, 빵, 초콜릿, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 기능수, 드링크제, 알코올음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다. 본 발명의 식품 조성물에서 발효 청미래덩굴 잎 추출물 또는 이의 분획물의 함량은 조성물 총 중량을 기준으로 0.01~50 중량%, 바람직하게는 0.1~25 중량%, 더 바람직하게는 0.5~10 중량%이나, 이에 한정되는 것은 아니다. 본 발명의 식품 조성물은 발효 청미래덩굴 잎 추출물 또는 이의 분획물 외에 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 또한, 본 발명의 식품 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 본 발명의 식품 조성물은 천연 과일주스, 과일주스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분들은 독립적으로 또는 혼합하여 사용할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 슈크로스와 같은 디사카라이드, 및 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드, 자일리톨, 소르비톨, 에리트리톨 등의 당알코올이다. 향미제로는 타우마틴, 스테비아 추출물과 같은 천연 향미제나 사카린, 아스파르탐과 같은 합성 향미제 등을 사용할 수 있다.
The food composition of the present invention comprising the fermented Cranberry Vine Leaf Extract or its fractions as an active ingredient may be in the form of pills, powders, granules, precipitates, tablets, capsules or liquid preparations. Examples of the food include meat, sausages, Beverage, tea, functional water, drink, alcoholic beverage, and vitamin complex, and the like, in general terms, such as bread, chocolate, candy, snack, confectionery, pizza, ramen and other noodles, gums, ice cream, All of the health foods in the. In the food composition of the present invention, the content of the fermented Cynthia rumulus leaf extract or its fraction is 0.01 to 50% by weight, preferably 0.1 to 25% by weight, more preferably 0.5 to 10% by weight based on the total weight of the composition, But is not limited thereto. The food composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient in addition to the fermented Cynthia raspberry extract or its fractions. In addition, the food composition of the present invention can be used as a food composition containing various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, , A carbonating agent used in carbonated drinks, and the like. In addition, the food composition of the present invention may contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The above-mentioned natural carbohydrates are sugar alcohols such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and xylitol, sorbitol and erythritol. Natural flavors such as tau Martin and stevia extract, and synthetic flavors such as saccharin and aspartame may be used as the flavor.
이하, 본 발명을 실시예를 통하여 보다 구체적으로 설명한다. 다만, 하기 실시예는 본 발명을 기술적 특징을 명확하게 예시하기 위한 것 일뿐, 본 발명의 보호범위를 한정하는 것은 아니다.
Hereinafter, the present invention will be described in more detail with reference to examples. However, the following examples are intended to clearly illustrate the technical features of the present invention and do not limit the scope of protection of the present invention.
Ⅰ. 제 1차 실험 : 청미래덩굴 잎 발효용 균주의 선별 및 발효방법 확립Ⅰ. The first experiment: Selection and fermentation method of strains for fermentation of Cheongmyeong vine leaf
1. 청미래덩굴 잎 분말의 준비1. Preparation of Chrysanthemum morifolium leaf powder
2012년 10월 대구광역시의 양령 시장에서 구입한 청미래덩굴(Smilax china L.) 잎을 수돗물로 깨끗이 세척한 후 열풍 건조기를 이용하여 40℃에서 충분히 건조시켰다. 이후 건조된 청미래덩굴(Smilax china L.) 잎을 분쇄기(모델명 : HNF-1710; 공급자 : Hanil Electric Co Ltd, Seoul, Korea)를 사용하여 약 40 메쉬(mesh) 정도의 입도 크기로 분쇄한 후 폴리에틸렌 필름으로 밀봉하여 4℃에서 보관하면서 후술하는 발효 시료로 사용하였다.
In October, 2012, the purchase of Smilax ( Smilax china L.) leaves were thoroughly cleaned with tap water and then sufficiently dried at 40 ° C using a hot air dryer. After the drying Smilax China (Smilax china L.) leaf was pulverized to a particle size of about 40 mesh using a pulverizer (model: HNF-1710; supplied by Hanil Electric Co Ltd, Seoul, Korea), sealed with a polyethylene film, And used as a fermentation sample described later.
2. 발효용 균주의 입수2. Acquisition of strain for fermentation
하기 표 1은 본 발명의 실험에서 사용한 균주의 종류를 나타낸 것이다. 하기 표 1 중 Aspergillus 속 미생물은 한국미생물배양센터(Korean Culture Center for Microorganisms, KCCM, Seoul, Korea)에서 구입하였으며 나머지 균주들은 한국식품연구원(Korea Food Research Institute, KFRI, Sungnam, Korea) 및 명지대학교 미생물학 교실에서 분양받았다.Table 1 below shows the types of strains used in the experiments of the present invention. Aspergillus The strains were purchased from the Korea Food Research Institute (KFRI, Sungnam, Korea) and the Myongji University microbiology class. The strains were purchased from the Korean Culture Center for Microorganisms (KCCM, Seoul, Korea).
* : 바실러스속 혼합 균주로는 전통청국장으로부터 분리한 바실러스 서쿨란스(B. circulans), 바실러스 브레비스(B. brevis), 바실러스 리케니포르미스(B. licheniformis), 바실러스 아라비탄(B. arabitane), 바실러스 코아굴란스(B. coagulans), 바실러스 스테아로써모필루스(B. stearothermophilus) 및 바실러스 서브틸리스(B. subtilis)의 혼합 균주를 사용하였다.*: The Bacillus sp. Strains include B. circulans , B. brevis , B. licheniformis , B. arabitane , Bacillus subtilis, Bacillus subtilis, Bacillus subtilis, A mixed strain of B. coagulans , B. stearothermophilus and B. subtilis was used.
** : 사카로마이세스 세레비지애(Saccharomyces cerevisiae)는 상업적으로 판매하고 있는 활성 건조효모(Premier Cuvee, Red Star Co, France)를 구입하여 사용하였다.
**: Saccharomyces cerevisiae was purchased from commercial commercial dried yeast (Premier Cuvee, Red Star Co, France).
3. 발효용 균주 배양액의 제조3. Preparation of culture broth for fermentation
(1) 아스퍼질러스 오리재(Aspergillus oryzae) 배양액의 준비(1) Aspergillus oryzae ) Preparation of culture medium
아스퍼질러스 오리재(Aspergillus oryzae) KACC 40247 균주를 Bacto™ malt extract(Becton, Dickinson and Company, Sparks, MD, USA)에 이식하고 25℃에서 10일 동안 진탕 배양하였다. 이후, 동일한 배지로 배양액 내 아스퍼질러스 오리재 농도를 108 cells/㎖로 조정하여 아스퍼질러스 오리재(Aspergillus oryzae) KACC 40247 배양액을 준비하였다. Aspergillus oryzae KACC 40247 strain was transplanted into Bacto ™ malt extract (Becton, Dickinson and Company, Sparks, MD, USA) and incubated with shaking at 25 ° C for 10 days. Then, by adjusting the culture medium within the Aspergillus duck material concentration in the same medium at 10 8 cells / ㎖ Aspergillus duck material (Aspergillus oryzae ) KACC 40247 culture was prepared.
Aspergillus oryzae KFRI 995 균주, Aspergillus oryzae var. brunneus KACC 44823 균주, Aspergillus oryzae var effuses KACC 44990 균주 각각에 대해서도 동일한 방법을 사용하여 발효용 균주 배양액을 준비하였다.
Aspergillus oryzae KFRI 995 strain, Aspergillus There are oryzae . brunneus KACC 44823 strain, Aspergillus oryzae var effuses For each of the KACC 44990 strains, a culture broth for fermentation was prepared using the same method.
(2) 바실러스속 혼합 균주 배양액의 준비(2) Preparation of Bacillus sp.
전통청국장으로부터 분리한 바실러스 서쿨란스(B. circulans), 바실러스 브레비스(B. brevis), 바실러스 리케니포르미스(B. licheniformis), 바실러스 아라비탄(B. arabitane), 바실러스 코아굴란스(B. coagulans), 바실러스 스테아로써모필루스(B. stearothermophilus) 및 바실러스 서브틸리스(B. subtilis)를 Bacto™ Tryptic Soy Broth(Becton, Dickinson and Co, Sparks, MD, USA)에 각각 이식하고 25℃에서 10일 동안 진탕 배양한 후 각각의 배양액을 동일 량으로 혼합하였다. 이후, 동일한 배지로 혼합 배양액 내 바실러스속(Bacillus) 미생물의 농도를 108 cells/㎖로 조정하여 바실러스속(Bacillus) 혼합 균주 배양액을 준비하였다.
B. circulans , B. brevis , B. licheniformis , B. arabitane , B. coagulans , and the like, which are separated from the conventional chungkukjang, ), as a brush loose Bacillus stearate (B. stearothermophilus) and Bacillus subtilis (B. subtilis), a Bacto ™ Tryptic Soy Broth (Becton, Dickinson and Co, Sparks, respectively implanted in the MD, USA) and from 25 ℃ 10 il After shaking culture, each culture was mixed in the same amount. Thereafter, the Bacillus microorganism concentration was adjusted to 10 8 cells / ml in the same culture medium to prepare a Bacillus mixed culture.
(3) 비피도박테리움 비피더스 배양액의 준비(3) Preparation of Bifidobacterium bifidus culture solution
비피도박테리움 비피더스(Bifidobacterium bifidus) ATCC 29521 균주를 Bacto™ MRS broth(Becton, Dickinson and Co, Sparks, MD, USA)에 이식하고 25℃에서 10일 동안 진탕 배양하였다. 이후, 동일한 배지로 배양액 내 비피도박테리움 비피더스 농도를 108 cells/㎖로 조정하여 비피도박테리움 비피더스(Bifidobacterium bifidus) 배양액을 준비하였다.
Bifidobacterium bifidus (Bifidobacterium bifidus strain ATCC 29521 was transplanted into Bacto ™ MRS broth (Becton, Dickinson and Co., Sparks, MD, USA) and incubated at 25 ° C for 10 days with shaking. Thereafter, the Bifidobacterium bifidus concentration in the culture medium was adjusted to 10 8 cells / ml with the same medium to obtain Bifidobacterium bifidus bifidus ) culture medium was prepared.
(4) 모나스커스 필로서스 배양액의 준비(4) Preparation of Monascus Lucillus Culture Medium
모나스커스 필로서스(Monascus pilosus) KCCM 60084 균주를 Bacto™ Tryptic Soy Broth(Becton, Dickinson and Co, Sparks, MD, USA)에 이식하고 25℃에서 10일 동안 진탕 배양하였다. 이후, 동일한 배지로 배양액 내 모나스커스 필로서스 농도를 108 cells/㎖로 조정하여 모나스커스 필로서스 배양액을 준비하였다.
Monascus pilosus ) KCCM 60084 strain was transplanted into BactoTM Tryptic Soy Broth (Becton, Dickinson and Co., Sparks, MD, USA) and cultured at 25 DEG C for 10 days with shaking. Thereafter, the concentration of Monascus purpilsus in the culture was adjusted to 10 8 cells / ml with the same medium to prepare a Monascus perfusos culture.
(5) 사카로마이세스 세레비지애 배양액의 준비(5) Preparation of Saccharomyces cerevisiae culture solution
상업적으로 판매하고 있는 활성 건조효모(Premier Cuvee, Red Star Co, France)를 Bacto™ malt extract(Becton, Dickinson and Company, Sparks, MD, USA)에 이식하고 25℃에서 10일 동안 진탕 배양하였다. 이후, 동일한 배지로 배양액 내 효모의 농도를 108 cells/㎖로 조정하여 사카로마이세스 세레비지애(Saccharomyces cerevisiae) 배양액을 준비하였다.
The commercially available active dry yeast (Premier Cuvee, Red Star Co, France) was implanted into Bacto ™ malt extract (Becton, Dickinson and Company, Sparks, MD, USA) and incubated at 25 ° C for 10 days with shaking. Thereafter, the yeast concentration in the culture medium was adjusted to 10 8 cells / ml with the same medium, and Saccharomyces cerevisiae cerevisiae were prepared.
4. 발효 청미래덩굴 잎의 제조4. Manufacture of Fermented Blue-faced Vine Leaves
앞에서 준비한 청미래덩굴 잎 분말 1.5㎏에 종균액과 물의 혼합액 3ℓ를 첨가하고 골고루 혼합하여 총 수분 함량을 약 67%가 되게 조정하였다. 이때, 종균액으로는 앞에서 준비한 발효용 균주 배양액을 사용하였다. 또한, 종균액과 물의 혼합액은 청미래덩굴 잎 분말 중량을 기준으로 5%(v/w)에 해당하는 종균액에 살균수를 혼합하여 총 3ℓ가 되도록 조정한 것이다. 이후, 청미래덩굴 잎 분말, 종균액 및 물의 혼합물을 플라스틱 발효 상자(가로 50㎝, 세로 70㎝, 높이 18㎝)에 약 5㎝의 두께로 담고 플라스틱 발효 상자의 상부를 폴리에틸렌 필름으로 덮은 후 약 30℃에서 10일간 발효시켰다. 발효가 완료된 청미래덩굴 잎을 폴리프로필렌 백(polypropylene bag)에 담고 오토클레이브(Autoclave) 안으로 옮겨 121℃에서 60분간 처리하여 살균하고, 40℃에서 건조하여 발효 청미래덩굴 잎을 제조하였다.
3 kg of the mixture of the seedling broth and water was added to 1.5 kg of the above-prepared Cheongmyeonghwa leaf powder, and the mixture was evenly mixed to adjust the total moisture content to about 67%. At this time, the culture solution for fermentation prepared above was used as the seed solution. In addition, the mixed solution of the seed solution and water is prepared by mixing the sterilized water to the seed solution corresponding to 5% (v / w) based on the weight of the seed powder of Cheongmaea vine leaf so that the total amount is 3 liters. Thereafter, a mixture of Cheongmyeonghwa leaf powder, a sterilized liquid and water was placed in a plastic fermentation box (50 cm in length, 70 cm in height, 18 cm in height) at a thickness of about 5 cm, the upper part of the plastic fermentation box was covered with a polyethylene film, Lt; 0 > C for 10 days. After fermentation, the leaves of Cheongmyeonghwa were placed in a polypropylene bag, transferred to an autoclave, sterilized by treatment at 121 ° C for 60 minutes, and dried at 40 ° C to produce fermented Cynomolgus cv.
5. 비발효 청미래덩굴 잎의 제조5. Manufacture of non-fermented blue-violet leaf
앞에서 준비한 청미래덩굴 잎 분말 1.5㎏에 물 3ℓ를 첨가하고 골고루 혼합하여 수분을 평형화시킨 후 이를 폴리프로필렌 백(polypropylene bag)에 담고 오토클레이브(Autoclave) 안으로 옮겨 121℃에서 60분간 처리하여 살균하였다. 이후, 살균된 청미래덩굴 잎 분말을 40℃에서 건조하여 비발효 청미래덩굴 잎을 제조하였다.
The water was equilibrated by adding 3 L of water to 1.5 Kg of the above-prepared CheongmyeRa vine leaf powder, and the water was equilibrated. The water was transferred into a polypropylene bag, transferred into an autoclave, and sterilized by treatment at 121 캜 for 60 minutes. Thereafter, the sterilized Cheongmyeongvin leaf powder was dried at 40 DEG C to prepare non-fermented blueberry leaf.
6. 발효 청미래덩굴 잎의 주요 기능성 지표 항목 측정 방법6. Main Functional Indicator Items of Fermented Blue-faced Vine Leaf Measuring Method
(1) 열수 추출액의 제조(1) Preparation of Hot Water Extract
차 추출기(Tea Extractor; Damian Tea Co, Anyang, Gyeonggi-Do, Korea)에 열수(약 90℃의 물)를 넣고 여기에 앞에서 제조한 발효 청미래덩굴 잎 또는 비발효 청미래덩굴 잎을 각각 열수 부피 대비 1 중량%(w/v)로 첨가한 후 5분간 우려내어 열수 추출액을 제조하였다. 제조한 열수 추출액을 후술하는 주요 기능성 지표 항목 측정의 시료로 사용하였다.
(About 90 ° C water) was added to a tea extractor (Damian Tea Co., Anyang, Gyeonggi-Do, Korea), and the fermented green tea leaves or non-fermented green tea leaves prepared above were added to 1 (W / v), followed by stirring for 5 minutes to prepare a hot water extract. The prepared hot water extract was used as a sample of the main functional index item measurement described later.
(2) 관능 검사에 의한 기호도 측정(2) Sensitivity measurement by sensory evaluation
추출액 시료를 약 50℃의 보온 통에 보관하면서 관능 검사용 시료로 사용하였다. 훈련된 관능 요원 25명을 대상으로 종합적인 기호도(overall acceptability)를 5점 측도법으로 평가하였다(Herbert & Jeol 1993). 5점 측도법의 평가 점수 기준은 아주 나쁘다 : 1점; 나쁘다 : 2점; 보통이다 : 3점; 좋다 : 4점; 아주 좋다 : 5점으로 하였다.
The extract samples were stored in a warm water bath at about 50 ° C and used as sensory test samples. The overall acceptability of 25 trained sensual agents was evaluated by the 5-point scoring method (Herbert & Jeol 1993). The five point scoring method scoring standard is very bad: 1 point; Bad: 2 points; Usually: 3 points; Good: 4 points; Very good: 5 points.
(3) 색상 측정(3) Color measurement
분광 광도계(spectrophotometer; Cary Winuv, Varian, Seoul, Korea)를 이용하여 추출액 시료에 대한 200~700㎚에서의 흡광도를 조사하고, 475㎚에서의 흡광도 값을 기준으로 색상을 비교하였다.
The absorbance at 200 to 700 nm of the extract samples was measured using a spectrophotometer (Cary Winuv, Varian, Seoul, Korea), and the color was compared based on the absorbance at 475 nm.
(4) 총 폴리페놀 함량 및 총 플라보노이드 함량 측정(4) Total polyphenol content and total flavonoid content
Minussi 등 (2003)의 방법에 따라 추출액 시료 100㎕에 2% 탄산나트륨(sodium carbonate) 용액 2 ㎖와 50% Folin-Ciacalteu reagent 100㎕를 가한 후 실온에서 약 1시간 동안 정치시키고, 720 ㎚에서 흡광도를 측정하였으며, 표준 물질인 갈산(gallic acid; Sigma-Aldrich Co, USA)의 표준 검량선에 의하여 총 폴리페놀(Total polyphenol, TP) 함량을 산출하였다. 또한, Meda 등 (2005)의 방법에 따라 추출액 시료 1 ㎖에 2% 염화알루미늄(aluminum chloride) 용액 1 ㎖와 50% 에탄올 수용액 1 ㎖를 첨가하고 혼합한 후 실온에서 10분간 반응시킨 다음 415 ㎚에서 흡광도를 측정하였으며, 표준 물질인 퀘르세틴(quracetin; Sigma-Aldrich Co, USA)의 표준 검량선에 의하여 총 플라보노이드(Total flavonoid, TF) 함량을 산출하였다.
2 μl of a 2% sodium carbonate solution and 100 μl of a 50% Folin-Ciacalteu reagent were added to 100 μl of the extract sample according to the method of Minussi et al. (2003), and the mixture was allowed to stand at room temperature for about 1 hour and absorbance at 720 nm And total polyphenol (TP) content was calculated by standard calibration curve of gallic acid (Sigma-Aldrich Co, USA). 1 ml of 2% aluminum chloride solution and 1 ml of 50% ethanol solution were added to 1 ml of the extract solution according to the method of Meda et al. (2005), and the mixture was reacted at room temperature for 10 minutes. Then, Absorbance was measured and total flavonoid (TF) content was calculated by standard calibration curve of quercetin (Sigma-Aldrich Co, USA).
(5) XOIA(Xanthine oxidase inhibitory activity) 및 AOIA(aldehyde oxidase inhibitory activity)의 측정(5) Measurement of XOIA (Xanthine oxidase inhibitory activity) and AOIA (aldehyde oxidase inhibitory activity)
XOIA(Xanthine oxidase inhibitory activity) 및 AOIA(aldehyde oxidase inhibitory activity)의 측정을 위한 효소원은 Rajagopalan 등(1962) 및 Maia 등(2002)의 방법에 따라 토끼의 간 조직으로부터 추출→황산암모늄(ammonium sulfate) 분획→투석→원심분리를 행하여 부분 정제한 후 -70℃에서 보관하면서 공시하였다. XOIA(Xanthine oxidase inhibitory activity)는 Stirpe & Della Corte(1969)의 방법에 따라 기질인 잔틴(xanthine)을 우릭산(uric acid)으로 전환하였을 때, 대조구(추출액 시료를 첨가하지 않은 것)와 비교하여 억제 정도를 %로 나타내었다. AOIA(aldehyde oxidase inhibitory activity)는 Rajagopalan 등(1962)의 방법에 따라 기질 NMN(N1-methylnicotinamide)으로부터 산화된 피리돈(pyridone)을 300 ㎚에서 측정한 다음 대조구(추출액 시료 대신 증류수를 첨가한 것)와 비교하여 억제 정도를 %로 나타내었다.
Enzymes for the measurement of XOIA and AOIA were extracted from liver tissue of rabbits according to the method of Rajagopalan et al. (1962) and Maia et al. (2002), and ammonium sulfate was used for the measurement of XOIA and aldehyde oxidase inhibitory activity. Fraction → dialysis → centrifugation, and then partially purified and stored at -70 ° C. Xanthine oxidase inhibitory activity (XOIA) was measured by the method of Stirpe & Della Corte (1969), when xanthine was converted to uric acid, compared with control (no extract sample) The degree of inhibition was expressed in%. The aldehyde oxidase inhibitory activity (AOIA) was determined by measuring the pyridone oxidized from the substrate NMN (N 1 -methylnicotinamide) at 300 nm according to the method of Rajagopalan et al. (1962), and then adding the distilled water ), The degree of inhibition was expressed in%.
7. 발효 청미래덩굴 잎의 주요 기능성 지표 항목 측정 결과7. Results of main functional index item measurement of fermented Cyanophyceae
하기 표 2에 발효 청미래덩굴 잎과 비발효 청미래덩굴 잎의 주요 기능성 지표 항목 측정 결과를 나타내었다.Table 2 shows the results of the measurement of the major functional index items of fermented blueberry leaf and nonfermented blueberry leaf.
표 2에서 보이는 바와 같이 아스퍼질러스속 미생물로 발효시킨 청미래덩굴 잎은 다른 미생물로 발효시킨 청미래덩굴 잎 또는 비발효 청미래덩굴 잎에 비해 플라보노이드의 흡수 파장으로 사료되는 475㎚ 파장에서의 피크 값이 높게 나타났다. 이러한 결과는 아스퍼질러스속 미생물에 의한 청미래덩굴 잎의 발효에 의하여 청미래덩굴 잎에 존재하는 폴리페놀 성분들이 플라보노이드 성분으로 변화됨과 동시에 발효에 의하여 추출 수율이 높아진 결과로 판단된다. 총 플라보노이드 함량 : 총 폴리페놀 함량의 비도 아스퍼질러스속 미생물로 발효시킨 청미래덩굴 잎에서 다른 미생물로 발효시킨 청미래덩굴 잎 또는 비발효 청미래덩굴 잎보다 높게 나타나 플라보노이드의 비율이 높은 것으로 확인되었다. 또한, 아스퍼질러스속 미생물로 발효시킨 청미래덩굴 잎은 다른 미생물로 발효시킨 청미래덩굴 잎 또는 비발효 청미래덩굴 잎에 비해 관능 검사에 의한 종합적인 기호도가 현저하게 우수한 것으로 평가되었다. 이러한 결과는 아스퍼질러스속 미생물에 의한 발효 과정에서 청미래덩굴 잎의 신맛이 중화되고 구감(Mouth feel)이 향상된 것에 기인한 것으로 판단된다. 또한, 아스퍼질러스속 미생물로 발효시킨 청미래덩굴 잎은 다른 미생물로 발효시킨 청미래덩굴 잎 또는 비발효 청미래덩굴 잎에 비해 XOIA(Xanthine oxidase inhibitory activity) 및 AOIA(aldehyde oxidase inhibitory activity)가 매우 높은 것으로 나타났고 그 차이는 AOIA(aldehyde oxidase inhibitory activity)에서 현저하였다. XO는 체내에서 ROS를 생성할 뿐만 아니라 요통의 원인물질인 요산을 생성하는 효소이며, AO 역시 ROS를 생성함과 동시에 알코올성 간 손상의 유도 및 고지방 식이에 의한 지방간 생성에 깊이 연관되어 있는 효소로 알려져 있다.
As shown in Table 2, the cheongmaea vine leaf fermented with aspergillus microorganisms had a peak value at a wavelength of 475 nm, which is considered to be the absorption wavelength of flavonoid, compared with the cheongmyeong vine leaf or the nonfermented cheongmyeong vine leaf fermented with other microorganisms . These results suggest that the fermentation of Aspergillus microorganisms resulted in the fermentation of polyphenols, which are present in the leaves of Cheongpyongjae, as a flavonoid component, resulting in higher yields. Total flavonoid content: The total polyphenol content was higher than that of non - fermented bluefin tuna or non fermented bluefin tuna vine leaves fermented with other microorganisms in non - asphalted microorganism fermented with the ratio of flavonoids. In addition, it was evaluated that Cheongmaea vine leaves fermented with aspergillus microorganisms were significantly superior to those of Cheongmyeong vine leaves or nonfermented Cheongmyeong vine leaves fermented with other microorganisms by sensory evaluation. These results suggest that the fermentation process of aspergillus microorganisms neutralizes the sourness of the leaves and improves the mouth feel. In addition, Xingtae oxidase inhibitory activity (XOIA) and aldehyde oxidase inhibitory activity (AOIA) were found to be higher than that of non-fermented Cynomolgus sinensis or Vinegar fermented by other microorganisms, as fermented by Aspergillus microorganism The difference was significant in AOIA (aldehyde oxidase inhibitory activity). XO is an enzyme that not only produces ROS in the body but also produces uric acid which is the cause of back pain. AO is also known to be involved in the induction of alcoholic liver damage and fatty liver formation by high fat diet as well as producing ROS have.
Ⅱ. 제 2차 실험 : 아스퍼질러스속 미생물에 의한 청미래덩굴 잎의 적정 발효기간 확립Ⅱ. Second Experiment: Establishment of optimum fermentation period of Aspergillus spp.
1. 청미래덩굴 잎 분말의 준비1. Preparation of Chrysanthemum morifolium leaf powder
2012년 10월 대구광역시의 양령 시장에서 구입한 청미래덩굴(Smilax china L.) 잎을 수돗물로 깨끗이 세척한 후 열풍 건조기를 이용하여 40℃에서 충분히 건조시켰다. 이후 건조된 청미래덩굴(Smilax china L.) 잎을 분쇄기(모델명 : HNF-1710; 공급자 : Hanil Electric Co Ltd, Seoul, Korea)를 사용하여 약 40 메쉬(mesh) 정도의 입도 크기로 분쇄한 후 폴리에틸렌 필름으로 밀봉하여 4℃에서 보관하면서 후술하는 발효 시료로 사용하였다.
In October, 2012, the purchase of Smilax ( Smilax china L.) leaves were thoroughly cleaned with tap water and then sufficiently dried at 40 ° C using a hot air dryer. After the drying Smilax China (Smilax china L.) leaf was pulverized to a particle size of about 40 mesh using a pulverizer (model: HNF-1710; supplied by Hanil Electric Co Ltd, Seoul, Korea), sealed with a polyethylene film, And used as a fermentation sample described later.
2. 발효용 균주 배양액의 제조2. Preparation of culture broth for fermentation
발효용 균주로는 이전의 실험에서 청미래덩굴 잎 발효용으로 가장 적합한 것으로 나타난 Aspergillus oryzae KFRI 995 균주를 사용하였다. As a strain for fermentation, the Aspergillus strain oryzae KFRI 995 strain was used.
Aspergillus oryzae KFRI 995 균주를 Bacto™ malt extract(Becton, Dickinson and Company, Sparks, MD, USA)에 이식하고 25℃에서 10일 동안 진탕 배양하였다. 이후, 동일한 배지로 배양액 내 아스퍼질러스 오리재 농도를 108 cells/㎖로 조정하여 Aspergillus oryzae KFRI 995 배양액을 준비하였다.
Aspergillus oryzae KFRI 995 strain was transplanted into Bacto (TM) malt extract (Becton, Dickinson and Company, Sparks, MD, USA) and cultured at 25 ° C for 10 days with shaking. Then, the concentration of aspergillus oryzae in the culture medium was adjusted to 10 8 cells / ml with the same medium, and Aspergillus oryzae KFRI 995 culture medium was prepared.
3. 발효 청미래덩굴 잎의 제조3. Preparation of Fermented Blue-faced Vine Leaves
앞에서 준비한 청미래덩굴 잎 분말 1.5㎏에 종균액과 물의 혼합액 3ℓ를 첨가하고 골고루 혼합하여 총 수분 함량을 약 67%가 되게 조정하였다. 이때, 종균액으로는 앞에서 준비한 발효용 균주 배양액을 사용하였다. 또한, 종균액과 물의 혼합액은 청미래덩굴 잎 분말 중량을 기준으로 5%(v/w)에 해당하는 종균액에 살균수를 혼합하여 총 3ℓ가 되도록 조정한 것이다. 이후, 청미래덩굴 잎 분말, 종균액 및 물의 혼합물을 플라스틱 발효 상자(가로 50㎝, 세로 70㎝, 높이 18㎝)에 약 5㎝의 두께로 담고 플라스틱 발효 상자의 상부를 폴리에틸렌 필름으로 덮은 후 약 30℃에서 각각 10일, 20일 및 30일간 발효시켰다. 발효 시간별로 발효가 완료된 청미래덩굴 잎을 폴리프로필렌 백(polypropylene bag)에 담고 오토클레이브(Autoclave) 안으로 옮겨 121℃에서 60분간 처리하여 살균하고, 40℃에서 건조하여 발효 청미래덩굴 잎을 제조하였다.
3 kg of the mixture of the seedling broth and water was added to 1.5 kg of the above-prepared Cheongmyeonghwa leaf powder, and the mixture was evenly mixed to adjust the total moisture content to about 67%. At this time, the culture solution for fermentation prepared above was used as the seed solution. In addition, the mixed solution of the seed solution and water is prepared by mixing the sterilized water to the seed solution corresponding to 5% (v / w) based on the weight of the seed powder of Cheongmaea vine leaf so that the total amount is 3 liters. Thereafter, a mixture of Cheongmyeonghwa leaf powder, a sterilized liquid and water was placed in a plastic fermentation box (50 cm in length, 70 cm in height, 18 cm in height) at a thickness of about 5 cm, the upper part of the plastic fermentation box was covered with a polyethylene film, Lt; 0 > C for 10 days, 20 days and 30 days, respectively. After fermentation, fermented green tea leaves were placed in a polypropylene bag, transferred into an autoclave, sterilized by treatment at 121 ° C for 60 minutes, and dried at 40 ° C to produce fermented Cyanophyceae.
4. 비발효 청미래덩굴 잎의 제조4. Manufacture of non-fermented blue-violet leaf
앞에서 준비한 청미래덩굴 잎 분말 1.5㎏에 물 3ℓ를 첨가하고 골고루 혼합하여 수분을 평형화시킨 후 이를 폴리프로필렌 백(polypropylene bag)에 담고 오토클레이브(Autoclave) 안으로 옮겨 121℃에서 60분간 처리하여 살균하였다. 이후, 살균된 청미래덩굴 잎 분말을 40℃에서 건조하여 비발효 청미래덩굴 잎을 제조하였다.
The water was equilibrated by adding 3 L of water to 1.5 Kg of the above-prepared CheongmyeRa vine leaf powder, and the water was equilibrated. The water was transferred into a polypropylene bag, transferred into an autoclave, and sterilized by treatment at 121 캜 for 60 minutes. Thereafter, the sterilized Cheongmyeongvin leaf powder was dried at 40 DEG C to prepare non-fermented blueberry leaf.
5. 발효 청미래덩굴 잎의 주요 기능성 지표 항목 측정 방법5. Main Functional Indicator Items of Fermented Blue-faced Vine Leaf Measuring Method
(1) 열수 추출액의 제조(1) Preparation of Hot Water Extract
차 추출기(Tea Extractor; Damian Tea Co, Anyang, Gyeonggi-Do, Korea)에 열수(약 90℃의 물)를 넣고 여기에 앞에서 제조한 발효 청미래덩굴 잎 또는 비발효 청미래덩굴 잎을 각각 열수 부피 대비 1 중량%(w/v)로 첨가한 후 5분간 우려내어 열수 추출액을 제조하였다. 제조한 열수 추출액을 후술하는 주요 기능성 지표 항목 측정의 시료로 사용하였다.
(About 90 ° C water) was added to a tea extractor (Damian Tea Co., Anyang, Gyeonggi-Do, Korea), and the fermented green tea leaves or non-fermented green tea leaves prepared above were added to 1 (W / v), followed by stirring for 5 minutes to prepare a hot water extract. The prepared hot water extract was used as a sample of the main functional index item measurement described later.
(2) 관능 검사(2) Sensory evaluation
추출액 시료를 약 50℃의 보온 통에 보관하면서 관능 검사용 시료로 사용하였다. 훈련된 관능 요원 25명을 대상으로 향(aroma), 맛(taste), 색상(color), 밝기(brightness), 입에 닿는 감각(mouth feel) 및 종합적인 기호도(overall acceptability)를 5점 측도법으로 평가하였다(Herbert & Jeol 1993). 5점 측도법의 평가 점수 기준은 아주 나쁘다 : 1점; 나쁘다 : 2점; 보통이다 : 3점; 좋다 : 4점; 아주 좋다 : 5점으로 하였다.
The extract samples were stored in a warm water bath at about 50 ° C and used as sensory test samples. The aroma, taste, color, brightness, mouth feel, and overall acceptability of 25 trained sensual agents were measured using a five point scoring method (Herbert & Jeol 1993). The five point scoring method scoring standard is very bad: 1 point; Bad: 2 points; Usually: 3 points; Good: 4 points; Very good: 5 points.
(3) 총 폴리페놀 함량 및 총 플라보노이드 함량 측정(3) Total polyphenol content and total flavonoid content
Minussi 등 (2003)의 방법에 따라 추출액 시료 100㎕에 2% 탄산나트륨(sodium carbonate) 용액 2 ㎖와 50% Folin-Ciacalteu reagent 100㎕를 가한 후 실온에서 약 1시간 동안 정치시키고, 720 ㎚에서 흡광도를 측정하였으며, 표준 물질인 갈산(gallic acid; Sigma-Aldrich Co, USA)의 표준 검량선에 의하여 총 폴리페놀(Total polyphenol, TP) 함량을 산출하였다. 또한, Meda 등 (2005)의 방법에 따라 추출액 시료 1 ㎖에 2% 염화알루미늄(aluminum chloride) 용액 1 ㎖와 50% 에탄올 수용액 1 ㎖를 첨가하고 혼합한 후 실온에서 10분간 반응시킨 다음 415 ㎚에서 흡광도를 측정하였으며, 표준 물질인 퀘르세틴(quracetin; Sigma-Aldrich Co, USA)의 표준 검량선에 의하여 총 플라보노이드(Total flavonoid, TF) 함량을 산출하였다.
2 μl of a 2% sodium carbonate solution and 100 μl of a 50% Folin-Ciacalteu reagent were added to 100 μl of the extract sample according to the method of Minussi et al. (2003), and the mixture was allowed to stand at room temperature for about 1 hour and absorbance at 720 nm And total polyphenol (TP) content was calculated by standard calibration curve of gallic acid (Sigma-Aldrich Co, USA). 1 ml of 2% aluminum chloride solution and 1 ml of 50% ethanol solution were added to 1 ml of the extract solution according to the method of Meda et al. (2005), and the mixture was reacted at room temperature for 10 minutes. Then, Absorbance was measured and total flavonoid (TF) content was calculated by standard calibration curve of quercetin (Sigma-Aldrich Co, USA).
(4) 전자공여능(Electron donating ability) 측정(4) Electron donating ability measurement
Blois(1958)의 방법에 따라 DPPH(1,1-diphenyl-2-picryl-hydrazyl) 어세이를 이용하여 전자공여능(Electron donating ability, 이하 EDA로 표기함)을 측정하였다. 추출액 시료 0.2㎖에 0.4mM DPPH(1,1-diphenyl-2-picryl-hydrazyl) 용액 0.8㎖를 가하고 10분간 방치한 다음, 525 ㎚에서 흡광도를 측정하였으며, 전자공여능은 다음과 같은 식으로 계산하였다. Electron donating ability (hereinafter referred to as EDA) was measured using a DPPH (1,1-diphenyl-2-picryl-hydrazyl) assay according to the method of Blois (1958). 0.8 ml of 0.4 mM DPPH (1,1-diphenyl-2-picryl-hydrazyl) solution was added to 0.2 ml of the extract sample, and the solution was allowed to stand for 10 minutes. Then, the absorbance at 525 nm was measured and the electron donating ability was calculated by the following equation .
EDA(%) = (1-A1/A0)×100EDA (%) = (1-A 1 / A 0 ) x 100
* A1 : 시료 첨가구의 흡광도* A 1 : Absorbance of sample added sphere
* A0 : 시료 무첨가구의 흡광도
* A 0 : Absorbance of sample without added sample
(5) 철 환원력(Ferric reducing antioxidant power, FRAP) 측정(5) Ferric reducing antioxidant power (FRAP) measurement
Benzie & Strani(1996)의 방법에 따라 추출액 시료 0.05 ㎖에 증류수 0.05 ㎖를 가하고 혼합한 후, 여기에 0.3M 아세트산 나트륨 버퍼(sodium acetate buffer, pH 3.60), 10 mM 2,4,6-tris (2-pyridyl)-s-triazine을 함유하는 40 mM HCl 용액 및 20 mM FeCl3 용액을 10:1:1의 부피 비로 혼합한 혼합용액 1.90 ㎖를 가하고 37℃에서 30분간 반응시킨 후 593 ㎚에서 흡광도를 측정하였다. FRAP(Ferric reducing antioxidant power)는 추출액 시료 제조를 위해 사용된 추출원료 건조 무게(g) 당 Fe3 +이 Fe2 +로 환원된 량을 μmole로 나타내었다.
0.05 ml of distilled water was added to 0.05 ml of the extract sample according to the method of Benzie & Strani (1996), mixed with 0.3 M sodium acetate buffer (pH 3.60), 10 mM 2,4,6-tris 2-pyridyl) -s-triazine and 20 mM FeCl 3 in a volume ratio of 10: 1: 1 was added, and the mixture was reacted at 37 ° C. for 30 minutes. Then, the absorbance at 593 nm Were measured. Ferric reducing antioxidant power (FRAP) is expressed in micromoles as the amount of Fe 3 + reduced to Fe 2 + per dry weight (g) of the extractive raw material used for the extraction sample preparation.
(6) 지질과산화 억제활성(Lipid peroxidation inhibitory activity, LPOIA) 측정(6) Measurement of Lipid peroxidation inhibitory activity (LPOIA)
지질과산화 억제활성(LPOIA)은 Banerjee 등(2005)의 방법을 일부 변형하여 측정하였다. 신선한 계란 노른자 10%(w/v)를 함유하는 50 mM Na-phosphate buffer(pH 7.4) 0.25㎖에 추출액 시료 0.20㎖와 증류수 0.55㎖을 가하였다. 여기에 70 mM FeSO4 용액 0.05㎖을 가하고 37℃에서 30분간 반응시킨 후, pH를 3.5로 조정한 20% 아세트산(acetic acid) 용액 1.5㎖, 0.8% TBA(thiobabituric acid)를 함유하는 도데실황산나트륨(sodium dodecyl sulfate) 1.5㎖ 및 20% 트리클로로아세트산(trichloroacetic acid) 0.05 ㎖를 가하고 95℃에서 1시간 동안 가열한 다음 냉각하였다. 여기에 n-부탄올(n-butanol) 5㎖를 가하고 3,000 rpm으로 10분간 원심분리하여 얻은 상층액의 흡광도를 532 ㎚에서 측정하였으며, 지질과산화 억제활성(LPOIA)은 다음과 같은 식으로 계산하였다.Lipid peroxidation inhibitory activity (LPOIA) was measured by modifying the method of Banerjee et al. (2005). 0.20 ml of extract sample and 0.55 ml of distilled water were added to 0.25 ml of 50 mM Na-phosphate buffer (pH 7.4) containing
LPOIA(%) = (1-A1/A0)×100LPOIA (%) = (1-A 1 / A 0 ) x 100
* A1 : 시료 첨가구의 흡광도* A 1 : Absorbance of sample added sphere
* A0 : 시료 무첨가구의 흡광도
* A 0 : Absorbance of sample without added sample
(7) XOIA(Xanthine oxidase inhibitory activity) 및 AOIA(aldehyde oxidase inhibitory activity)의 측정(7) Measurement of XOIA (Xanthine oxidase inhibitory activity) and AOIA (aldehyde oxidase inhibitory activity)
XOIA(Xanthine oxidase inhibitory activity) 및 AOIA(aldehyde oxidase inhibitory activity)의 측정을 위한 효소원은 Rajagopalan 등(1962) 및 Maia 등(2002)의 방법에 따라 토끼의 간 조직으로부터 추출→황산암모늄(ammonium sulfate) 분획→투석→원심분리를 행하여 부분 정제한 후 -70℃에서 보관하면서 공시하였다. XOIA(Xanthine oxidase inhibitory activity)는 Stirpe & Della Corte(1969)의 방법에 따라 기질인 잔틴(xanthine)을 우릭산(uric acid)으로 전환하였을 때, 대조구(추출액 시료를 첨가하지 않은 것)와 비교하여 억제 정도를 %로 나타내었다. AOIA(aldehyde oxidase inhibitory activity)는 Rajagopalan 등(1962)의 방법에 따라 기질 NMN(N1-methylnicotinamide)으로부터 산화된 피리돈(pyridone)을 300 ㎚에서 측정한 다음 대조구(추출액 시료 대신 증류수를 첨가한 것)와 비교하여 억제 정도를 %로 나타내었다.
Enzymes for the measurement of XOIA and AOIA were extracted from liver tissue of rabbits according to the method of Rajagopalan et al. (1962) and Maia et al. (2002), and ammonium sulfate was used for the measurement of XOIA and aldehyde oxidase inhibitory activity. Fraction → dialysis → centrifugation, and then partially purified and stored at -70 ° C. Xanthine oxidase inhibitory activity (XOIA) was measured by the method of Stirpe & Della Corte (1969), when xanthine was converted to uric acid, compared with control (no extract sample) The degree of inhibition was expressed in%. The aldehyde oxidase inhibitory activity (AOIA) was determined by measuring the pyridone oxidized from the substrate NMN (N 1 -methylnicotinamide) at 300 nm according to the method of Rajagopalan et al. (1962), and then adding the distilled water ), The degree of inhibition was expressed in%.
6. 발효 청미래덩굴 잎의 주요 기능성 지표 항목 측정 결과6. Results of main functional index item measurement of fermented blueberry leaf
Aspergillus oryzae KFRI 995로 청미래덩굴 잎을 발효하여 얻은 발효 청미래덩굴 잎의 발효 시간에 따른 주요 기능성 지표 항목 측정 결과에서 "NF"는 비발효 청미래덩굴 잎의 열수 추출액을, "F10"은 10일 동안 발효하여 얻은 발효 청미래덩굴 잎의 열수 추출액을, "F20"은 20일 동안 발효하여 얻은 발효 청미래덩굴 잎의 열수 추출액을, "F30"은 30일 동안 발효하여 얻은 발효 청미래덩굴 잎의 열수 추출액을 각각 나타낸다.
Aspergillus oryzae KFRI 995, "NF" refers to the fermentation of non-fermented blueberry leaf vine leaf, "F10" refers to fermentation for 10 days, "F20" represents the hot water extract of the fermented Cyanophyceae obtained by fermentation for 20 days, and "F30" represents the hot water extract of the fermented Cyanophyceae obtained by fermentation for 30 days, respectively.
(1) 관능 검사 결과(1) Sensory test results
하기 표 3은 Aspergillus oryzae KFRI 995로 청미래덩굴 잎을 발효하여 얻은 발효 청미래덩굴 잎의 발효 시간에 따른 관능 검사 결과를 나타낸 것이다. 하기 표 3에서 보이는 바와 같이 향에 대한 기호도는 비발효 청미래덩굴 잎 및 발효 청미래덩굴 잎 모두에서 유의적인 차이를 보이지 않았다. 맛에 대한 기호도는 10일 내지 30일간 발효시킨 F10, F20, F30 상호 간에는 유의적인 차이를 보이지 않았으나 발효시킨 경우가 발효시키지 않은 NF에 비하여는 양호한 것으로 나타났다. 이러한 현상은 발효시키지 않은 청미래덩굴 잎은 비교적 강한 신맛과 떫은맛을 띠는데 비하여 발효시킨 경우는 이러한 맛이 없거나 감소하고 이들 맛에 덮여 있던 감미가 비교적 뚜렷하게 나타난 때문이라 사료된다. 차의 밝기에 대한 기호도는 NF, F10, F20 및 F30 간에 유의적인 차이가 없었다. NF의 경우는 연한 황색을 띠는 반면 F10, F20 및 F30은 맑은 적색을 띠어 발효에 의하여 밝기에 대한 기호도가 낮아질 것으로 예상한 바와는 다른 결과를 나타내었다. 이러한 결과는 발효에 의하여 청미래덩굴 잎에 존재하는 청색을 띠는 엽록소(chlorophyll)이 소실되면서 F10, F20 및 F30에 해당하는 열수 추출액이 맑고 깨끗해 보이는 색상으로 변화된 때문으로 판단된다. 또한, 입에 닿는 감각과 종합적인 기호도도 맛에 대한 기호도와 동일하게 발효시킨 청미래덩굴 잎이 발효시키지 않은 청미래덩굴 잎에 비하여 유의적으로 높은 값을 나타내었다. 향(aroma), 맛(taste), 밝기 (brightness) 및 입에 닿는 감각 (mouth feel)에 대한 기호도는 특히 발효 차의 주요 품질평가 항목으로 알려져 있다(Owuor 등 2006; Chen 등 2000). 일반적으로 생엽에 존재하는 엽록소가 발효에 의하여 분해되어 소실되지 않으면 이들이 어두운 색상을 띠게 되어 밝기에 대한 기호도를 떨어트리게 된다. 또한, 차 잎 발효의 중요한 특징은 차 잎의 주요 성분인 카테킨(catechin)을 비롯한 폴리페놀(polyphenol) 성분을 폴리페놀 옥시다아제(polyphenol oxidase)에 의하여 산화, 중합시킴으로써 새로운 기능성 성분인 차 플라빈(tea flavins) 또는 차 루비진(tea rubigins)을 생성하고 이로 인해 적색계의 아름다운 색상을 띄게 함과 동시에 부드러운 맛을 갖게 하고 소화성을 향상시키는데에 있다(Lee 등 2012). Table 3 < RTI ID = 0.0 > oryzae KFRI 995 showed the sensory evaluation according to fermentation time. As shown in Table 3 below, the degree of preference for flavor did not show a significant difference in both non-fermented blue-violet creeper blade and fermented blueberry vine leaf. The preference for flavor did not show any significant difference between F10, F20 and F30 fermented for 10 to 30 days, but fermentation was better than non - fermented NF. This phenomenon is due to the relatively strong acidity and pungent taste of non - fermented Cynomolgus sinensis leaf, whereas when fermented, the flavor was reduced or decreased and the sweetness covered in these tastes was relatively clear. There was no significant difference between the NF, F10, F20 and F30 for the brightness of the car. NF showed light yellowish color while F10, F20 and F30 had a clear red color and showed different results from those expected to lower the brightness by fermentation. These results indicate that the fermentation resulted in the disappearance of the blue chlorophyll present on the leaves of the bluegrass, and that the hot water extracts corresponding to F10, F20 and F30 changed to a clear and clean color. In addition, sensory touch and overall preference were significantly higher than those of non - fermented Cheongpyonga vine leaves which were fermented in the same manner as taste preference. The aroma, taste, brightness and taste of the mouth are known to be the main quality evaluation items of fermented tea (Owuor et al. 2006; Chen et al. 2000). Generally, if the chlorophyll present in the green leaf is decomposed by fermentation and does not disappear, they become darker colors, which reduces the preference for brightness. In addition, an important feature of tea leaf fermentation is the oxidation and polymerization of polyphenol components including catechin, which is a main component of tea leaves, by polyphenol oxidase, flavins, or tea rubigins, thereby providing a beautiful color of the red color system, at the same time having a soft taste and improving the digestibility (Lee et al., 2012).
(2) 총 폴리페놀(TP) 함량 및 총 플라보노이드(TF) 함량 측정 결과 (2) Total polyphenol (TP) content and total flavonoid (TF) content
도 1은 Aspergillus oryzae KFRI 995로 청미래덩굴 잎을 발효하여 얻은 발효 청미래덩굴 잎의 발효 시간에 따른 총 폴리페놀(TP) 함량 및 총 플라보노이드(TF) 함량 측정 결과를 나타낸 그래프이다. 도 1에서 보이는 바와 같이 발효 시간이 길어짐에 따라 TP 함량 및 TF의 함량이 점진적으로 감소함을 알 수 있다. 그러나, 발효 시간에 따라 TP의 감소율이 TF의 감소율에 비하여 상대적으로 커서 F20과 F30에서 TF:TP의 비율(%)은 각각 97.76% 및 89.38%로 나타났고, 이는 폴리페놀(polyphenol)의 대부분이 플라보노이드(flavonoid)임을 의미한다. 발효에 의한 폴리페놀(polyphenol) 성분의 감소는 발효도, 발효기간 및 발효방법에 따라 차이를 보이는 것으로 알려져 있으며 발효시에 비수용성 폴리페놀(polyphenol) 성분들이 수용성의 플라보노이드(flavonoid)로 전환한다는 보고도 있다 (Lee 등 2012).Figure 1 shows the Aspergillus (TP) content and total flavonoid (TF) content of Fermented Cynomolgus sinensis leaf obtained by fermentation of Cheongmyeongvle leaf with oryzae KFRI 995 according to fermentation time. As shown in FIG. 1, it can be seen that the TP content and the TF content gradually decrease as the fermentation time becomes longer. However, the decrease rate of TP was relatively higher than that of TF according to the fermentation time, and the ratio (%) of TF: TP in F20 and F30 was 97.76% and 89.38%, respectively. This indicates that most of the polyphenol It is a flavonoid. The reduction of polyphenol content by fermentation is known to vary depending on the fermentation degree, fermentation period and fermentation method, and it is reported that the nonpolyphenol polyphenols are converted into water soluble flavonoids during fermentation (Lee et al. 2012).
전통 메주발효균인 아스퍼질러스 오리재(Aspergillus oryzae)를 포함한 아스퍼질러스속 미생물은 발효시 아밀라아제(amylase), 프로테아제(protease)와 같은 효소를 강력하게 분비할 뿐만 아니라 펙티나아제(pectinase)와 같은 식물체 세포벽 성분의 분해에 관여하는 효소를 분비함으로써 세포벽과 강하게 결합하여 용출되기 어려운 기능성 성분들을 쉽게 가용화되게 하는 효과가 있다(Angayarkanni 등 2002). 또한, 아스퍼질러스속 미생물은 발효시 탄나아제(tannase)를 분비함으로써(Garcia-Conesa 등 2001) 식물체에 널리 분포하는 떫은맛의 원인이 되는 갈로탄닌(galloannins) 등을 분해시켜 떫은맛을 감소시키고 폴리페놀 함량을 감소시키는 효과가 있다. 식물체에 존재하는 폴리페놀(polyphenol) 성분은 항산화 작용을 비롯한 다양한 생리활성을 나타내며(Osawa 1994), 그 함량에 비례하여 항산화력이 높아지는 것으로 보고되고 있다(Holasova 등 2002). 그러나 한편으로는 폴리페놀(polyphenol) 성분의 함량이 높을수록 활성산소를 생성하는 과산화수소의 함량이 높다는 보고가 있으며(Arakawa 등 2004), 또한 폴리페놀 성분들은 소화작용을 억제하고 떫은맛을 나타내어 기호도를 떨어뜨리는 것으로 알려져 있다 (Lee 등 2012).
Aspergillus microorganisms including Aspergillus oryzae, which is a traditional meju fermenting microorganism, secrete not only enzymes such as amylase and protease during fermentation, but also plants such as pectinase By secretion of enzymes involved in the degradation of cell wall components, it has the effect of easily solubilizing functional components which are strongly bound to the cell wall and are difficult to elute (Angayarkanni et al. 2002). In addition, aspergillus microorganisms secrete tannase during fermentation (Garcia-Conesa et al. 2001), which degrades galloannins and the like, which are widely distributed in plants, There is an effect of reducing the content. The polyphenol component in plants has been reported to exhibit various physiological activities including antioxidant activity (Osawa 1994), and antioxidant activity is increased in proportion to its content (Holasova et al. 2002). On the other hand, it has been reported that the content of polyphenol is higher and the content of hydrogen peroxide producing active oxygen is higher (Arakawa 2004), and polyphenols are known to inhibit digestion and reduce the taste of pungent taste (Lee et al., 2012).
(3) 전자공여능(Electron donating ability, EDA) 및 철 환원력(Ferric reducing antioxidant power, FRAP) 측정 결과(3) Electron donating ability (EDA) and ferric reducing antioxidant power (FRAP)
도 2는 Aspergillus oryzae KFRI 995로 청미래덩굴 잎을 발효하여 얻은 발효 청미래덩굴 잎의 발효 시간에 따른 전자공여능(Electron donating ability, EDA) 측정 결과를 나타낸 그래프이고, 도 3은 Aspergillus oryzae KFRI 995로 청미래덩굴 잎을 발효하여 얻은 발효 청미래덩굴 잎의 발효 시간에 따른 철 환원력(Ferric reducing antioxidant power, FRAP) 측정 결과를 나타낸 그래프이다.FIG. 2 shows the Aspergillus FIG. 3 is a graph showing the results of measuring the electron donating ability (EDA) according to the fermentation time of the fermented blueberry leaf vine leaves obtained by fermenting Cheongmyeongvin leaf with oryzae KFRI 995, FIG. 3 is a graph showing the results of Aspergillus (ferric reducing antioxidant power, FRAP) according to the fermentation time of the fermented Cynomolgus sinensis leaf obtained by fermenting Cheongmyeongvin leaf with oryzae KFRI 995.
EDA는 체내에서 생성되는 ROS에 전자를 공여하여 산화를 막는 작용을 하며(Choi 등 2003), FRAP는 철 산화물을 원래의 상태로 환원시키는 힘을 의미한다. EDA와 FRAP는 모두 항산화를 나타내는 중요한 지표로 활용되며(Song 등 2006), EDA가 높을수록 FRAP도 높아지는 것으로 알려져 있다(Kang 등 1995). 도 2 내지 도 3에서 보이는 바와 같이 발효 시간이 길어짐에 따라 추출물 시료의 EDA 및 FRAP는 점진적으로 감소하였으나, 큰 차이를 보이지 않았다. Ko와 Yang(2011)은 청미래덩굴 잎 열수 추출 농축물의 EDA는 33.6%라 보고한바 있으며, 본 실험에서 비발효 청미래덩굴 잎의 1% 열수 추출액의 EDA는 29.01%로 이들의 결과와 유사하였다. 한편, EDA와 같은 항산화능은 폴리페놀(polyphenol)의 함량이 높을수록 높은 것으로 알려져 있으나(Torel 등 1986), 본 실험에서는 발효 시간이 늘어남에 따라 발효 청미래덩굴 잎의 폴리페놀(polyphenol)의 함량이 발효 10일째는 24.91%, 20일째는 56.92%, 30일째는 64.41%로 감소함에도 불구하고 EDA와 FRAP의 감소율은 폴리페놀(polyphenol)의 감소율에는 미지지 않을 정도로 비교적 낮았다. 이러한 결과는 아스퍼질러스속 미생물에 의한 청미래덩굴 잎의 발효에 의해 항산화능을 지니는 새로운 물질이 생성된 것에 기인한 것으로 판단된다. 다른 차 잎의 발효에서도 카테킨(catechin)과 같은 항산화성 물질이 감소하는 대신에 새로운 항산화 물질인 차 플라빈(teaflavins)나 차 루비진(tharubigins)가 생성되는 것으로 보고되고 있다 (Lee 등 2012).
EDA inhibits oxidation by donating electrons to the ROS produced in the body (Choi et al. 2003). FRAP means the power to reduce iron oxide to its original state. EDA and FRAP are both important indicators of antioxidant activity (Song et al. 2006). It is known that the higher the EDA, the higher the FRAP (Kang et al. 1995). As shown in FIGS. 2 to 3, EDA and FRAP of the extract samples gradually decreased with increasing fermentation time, but there was no significant difference. Ko and Yang (2011) reported 33.6% EDA of the hydrothermal extract of Cheongmyeongvle leaf. In this experiment, the EDA of 1% hydrothermal extract of nonfermented Cheonggyeong vine leaf was 29.01%, which is similar to the results. The antioxidant activity of EDA is known to be higher as the content of polyphenol is higher (Torel et al., 1986). However, as the fermentation time increases, the content of polyphenol in fermented blueberry leaves The reduction rate of EDA and FRAP was relatively low, which was not enough to observe the decreasing rate of polyphenol, even though it decreased to 24.91% on the 10th day of fermentation, 56.92% on the 20th day and 64.41% on the 30th day. These results are attributed to the production of new antioxidant substances by the fermentation of Aspergillus sp. It has also been reported that in the fermentation of other tea leaves, antioxidative substances such as catechin are reduced and new antioxidants such as teaflavins and tharubigins are produced (Lee et al., 2012).
(4) 지질과산화 억제활성(Lipid peroxidation inhibitory activity, LPOIA) 측정 결과(4) Lipid peroxidation inhibitory activity (LPOIA) measurement results
도 4는 Aspergillus oryzae KFRI 995로 청미래덩굴 잎을 발효하여 얻은 발효 청미래덩굴 잎의 발효 시간에 따른 지질과산화 억제활성(Lipid peroxidation inhibitory activity, LPOIA) 측정 결과를 나타낸 그래프이다.Figure 4 shows the Aspergillus (Lipid peroxidation inhibitory activity (LPOIA)) according to the fermentation time of fermented Cynomolgus sinensis leaf obtained by fermenting Cynomolgus sinensis leaf with oryzae KFRI 995.
도 4에서 보이는 바와 같이 발효 시간이 길어짐에 따라 추출물 시료의 LPOIA는 점진적으로 감소하였으나, 큰 차이를 보이지 않았다. LPOIA는 생체막에 존재하는 불포화지방산이 산화, 분해되어 독성을 나타내는 알데하이드(aldehyde), 케톤(ketone) 및 락톤(lactone)과 같은 물질이 생성(Hur 등 2004)되는 것을 차단하는 힘을 의미한다. 도 4에서 보이는 결과에 의할 때, 청미래덩굴 잎을 아스퍼질러스속 미생물로 발효하여도 LPOIA의 잔존 활성이 비교적 높게 유지되었고, 산화적 스트레스로 인한 세포조직의 손상을 경감시킬 수 있는 기능성 소재로서 발효 청미래덩굴 잎의 활용성이 기대된다.
As shown in FIG. 4, as the fermentation time became longer, the LPOIA of the extract samples gradually decreased, but there was no significant difference. LPOIA refers to the ability to block the formation of substances such as aldehyde, ketone, and lactone, which are toxic by oxidizing and degrading unsaturated fatty acids present in the biomembrane (Hur et al. 2004). 4, the residual activity of LPOIA remained relatively high even when fermented with Aspergillus sp. Microorganism, and as a functional material capable of alleviating cellular tissue damage due to oxidative stress, fermentation It is expected that the use of Cheongmaea vine leaves.
(5) XOIA(Xanthine oxidase inhibitory activity) 및 AOIA(aldehyde oxidase inhibitory activity)의 측정 결과(5) Measurement results of XOIA (Xanthine oxidase inhibitory activity) and AOIA (aldehyde oxidase inhibitory activity)
하기 표 4는 Aspergillus oryzae KFRI 995로 청미래덩굴 잎을 발효하여 얻은 발효 청미래덩굴 잎의 발효 시간에 따른 XOIA 및 AOIA 측정 결과를 나타낸 것이다.Table 4 below shows the Aspergillus The results of XOIA and AOIA measurement according to the fermentation time of the fermented Cynomolgus sinensis leaf obtained by fermenting the leaves of orangzae KFRI 995.
상기 표 4에서 보이는 바와 같이 비발효 청미래덩굴 잎과 발효 청미래덩굴 잎의 XOIA는 약 4~6%로 낮게 나타났으나, AOIA는 약 30~50%로 높게 나타났고, XOIA 및 AOIA는 발효에 의해 증가하는 것으로 나타났다. 이러한 결과는 청미래덩굴 뿌리인 토복령 추출물이 XO의 활성을 억제함으로서 통풍과 관련이 있는 요산의 생성을 억제한다는 보고(Chena 등 2011)와 차이를 보였으나, 발효 청미래덩굴 잎은 비교적 높은 AO 억제 활성을 보여 이와 관련되는 기능적인 효과가 크게 기대된다. XO와 AO는 동식물계에 널리 분포하며 종에 따라 특성적 차이를 나타내는 것으로 알려져 있다(Yasuhara 등 2002). XO와 AO는 기질 특이성에서는 약간의 차이를 보이지만 생화학적으로는 대단히 유사한 특성을 가진 효소(Yamaguchi 등 2007)로 피리미딘(pyrimidine), 퓨린(purine), 프테리딘(pteridine) 유도체와 같은 N-헤테로고리 화합물(Garattini 등 2008)이나 친유성 방향족 화합물의 산화에 관여한다 (Krenitsky 1978). 또한, 아조(azo) 염료(Stoddart & Levine, 1992), 항암제(Itoh 2009) 및 항고혈압제(Dambrova 등 1998)의 환원에 관여하여 때로는 생체 내에서 이들 약물의 효과를 감소시키기도 하지만 치료 후 체내에 잔존하는 약성을 해독하는 작용도 한다. AO는 XO가 결핍된 경우에 알로퓨리놀(allopurinol)과 피라진아마이드(pyrazinamide) 등의 대사에 관여하며(Moriwaki 등 1993), 두 효소 모두 분자 상의 산소를 전자수용체로 이용하여 슈퍼옥사이드(superoxide)와 과산화수소(hydrogen peroxide) 등과 같은 ROS을 생성하는 효소이다(Kundu 등 2007). 특히, AO는 에탄올에 의한 간 손상(Shaw와 Jayatilleke, 1990), 티오아세트아마이드(thioacetamide) 및 클로로포름(chloroform)에 의한 간 손상 (Ali 등 2008)을 촉진하고 지방세포(adipocyte)의 분화와 지질의 저장 및 지방세포 내의 단백질 (adiponectin) 분비에 관여하는 등 비만과 관련이 있는 것으로 보고되고 있다 (Johanna 등 2008). 또한, AO는 고지방식이 및 알코올성 지방간 생성에 관여(Shaw와 Jayatilleke 1990) 할 뿐만 아니라 항비만 아디포카인(adipokine)인 아디포넥틴(adiponectin)에 의해 AO의 활성이 저하된다는 보고도 있다 (Neumeier 등 2006). 일반적으로 식물체에 함유된 플라보노이드(flavonoid)는 AO의 활성을 억제하며 그 억제 정도는 플라보노이드(flavonoid)의 글리코시드(glycoside) 형태에 비하여 아글리콘(aglycone) 형태에서 현저하다 (Rashidi와 Nazemiyeh 2010). 본 실험에서 아스퍼질러스속 미생물에 의해 청미래 덩굴 잎이 발효되었을 때 TP와 TF의 함량이 감소함에도 AO 억제 활성은 오히려 증가하였으며, 이러한 결과는 발효에 의하여 청미래덩굴 잎에 존재하는 글리코시드(glycoside) 형태의 플라보노이드(flavonoid)가 아글리콘(aglycone) 형태로 분해됨을 시사한다.
As shown in Table 4, the XOIA of the non-fermented blueberry seedlings and the fermented blueberry seedlings was as low as about 4 to 6%, but the AOIA was as high as about 30 to 50%, and the XOIA and AOIA were fermented Respectively. These results were compared with those reported by Chena et al. (2011) that the extract of Toyohaku, which is a root of Cheongmaea japonica, inhibits the production of uric acid associated with gout by inhibiting the activity of XO. However, And the functional effect related to this is greatly expected. XO and AO are widely distributed in plant and animal systems and are known to exhibit characteristic differences depending on species (Yasuhara et al., 2002). XO and AO have very similar biochemical characteristics (Yamaguchi et al. 2007), showing a slight difference in the substrate specificity, but also in the case of pyrimidine, purine, and pteridine derivatives, It is involved in the oxidation of heterocyclic compounds (Garattini et al. 2008) and lipophilic aromatic compounds (Krenitsky 1978). In addition, azo dyes (Stoddart & Levine, 1992), anticancer drugs (Itoh 2009) and antihypertensive drugs (Dambrova 1998), and sometimes reduce the effects of these drugs in vivo, but they also have the function of detoxifying the drug remaining in the body after treatment. AO is involved in the metabolism of allopurinol and pyrazinamide in the absence of XO (Moriwaki et al. 1993). Both enzymes use oxygen as a molecular receptor to generate ROS such as superoxide and hydrogen peroxide (Kundu et al. (2007). In particular, AO promotes hepatic damage by ethanol (Shaw and Jayatilleke, 1990), thioacetamide and chloroform (Ali et al. 2008), and differentiation of adipocyte and lipid And is involved in the secretion of adiponectin in the adipocyte, and is associated with obesity (Johanna et al. 2008). It has also been reported that AO is involved in the production of high-fat diets and alcoholic fatty liver (Shaw and Jayatilleke 1990), as well as that the activity of AO is reduced by the adiponectin adiponectin (Neumeier et al. 2006). In general, flavonoids contained in plants inhibit the activity of AO, and the degree of inhibition is remarkable in the aglycone form compared to the glycoside form of flavonoid (Rashidi and Nazemiyeh 2010). In this experiment, AO inhibitory activity was increased even though TP and TF contents decreased when Aspergillus microflora fermented Cheongpyongjae leaves. These results showed that glycoside type Suggesting that the flavonoid of the plant is degraded into an aglycone form.
Ⅲ. 제 3차 실험 : 발효 청미래덩굴 잎의 Ⅲ. Tertiary Experiment: Fermentation of Chrysanthemum vine leaf
열수Heat number
추출물이 알코올 급여 마우스의 혈청 및 간 조직에 미치는 영향 Effect of Extracts on Serum and Liver Tissue of Alcohol-fed Mice
1. 청미래덩굴 잎 분말의 준비1. Preparation of Chrysanthemum morifolium leaf powder
2012년 10월 대구광역시의 양령 시장에서 구입한 청미래덩굴(Smilax china L.) 잎을 수돗물로 깨끗이 세척한 후 열풍 건조기를 이용하여 40℃에서 충분히 건조시켰다. 이후 건조된 청미래덩굴(Smilax china L.) 잎을 분쇄기(모델명 : HNF-1710; 공급자 : Hanil Electric Co Ltd, Seoul, Korea)를 사용하여 약 40 메쉬(mesh) 정도의 입도 크기로 분쇄한 후 폴리에틸렌 필름으로 밀봉하여 4℃에서 보관하면서 후술하는 발효 시료로 사용하였다.
In October, 2012, the purchase of Smilax ( Smilax china L.) leaves were thoroughly cleaned with tap water and then sufficiently dried at 40 ° C using a hot air dryer. After the drying Smilax China (Smilax china L.) leaf was pulverized to a particle size of about 40 mesh using a pulverizer (model: HNF-1710; supplied by Hanil Electric Co Ltd, Seoul, Korea), sealed with a polyethylene film, And used as a fermentation sample described later.
2. 발효용 균주 배양액의 제조2. Preparation of culture broth for fermentation
발효용 균주로는 이전의 실험에서 청미래덩굴 잎 발효용으로 가장 적합한 것으로 나타난 Aspergillus oryzae KFRI 995 균주를 사용하였다. As a strain for fermentation, the Aspergillus strain oryzae KFRI 995 strain was used.
Aspergillus oryzae KFRI 995 균주를 Bacto™ malt extract(Becton, Dickinson and Company, Sparks, MD, USA)에 이식하고 25℃에서 10일 동안 진탕 배양하였다. 이후, 동일한 배지로 배양액 내 아스퍼질러스 오리재 농도를 108 cells/㎖로 조정하여 Aspergillus oryzae KFRI 995 배양액을 준비하였다.
Aspergillus oryzae KFRI 995 strain was transplanted into Bacto (TM) malt extract (Becton, Dickinson and Company, Sparks, MD, USA) and cultured at 25 ° C for 10 days with shaking. Then, the concentration of aspergillus oryzae in the culture medium was adjusted to 10 8 cells / ml with the same medium, and Aspergillus oryzae KFRI 995 culture medium was prepared.
3. 발효 청미래덩굴 잎 및 이의 열수 추출물의 제조3. Preparation of fermented Cynomolgus vine leaf and its hot water extract
앞에서 준비한 청미래덩굴 잎 분말 1.5㎏에 종균액과 물의 혼합액 3ℓ를 첨가하고 골고루 혼합하여 총 수분 함량을 약 67%가 되게 조정하였다. 이때, 종균액으로는 앞에서 준비한 발효용 균주 배양액을 사용하였다. 또한, 종균액과 물의 혼합액은 청미래덩굴 잎 분말 중량을 기준으로 5%(v/w)에 해당하는 종균액에 살균수를 혼합하여 총 3ℓ가 되도록 조정한 것이다. 이후, 청미래덩굴 잎 분말, 종균액 및 물의 혼합물을 플라스틱 발효 상자(가로 50㎝, 세로 70㎝, 높이 18㎝)에 약 5㎝의 두께로 담고 플라스틱 발효 상자의 상부를 폴리에틸렌 필름으로 덮은 후 약 30℃에서 각각 10일, 20일 및 30일간 발효시켰다. 발효 시간별로 발효가 완료된 청미래덩굴 잎을 폴리프로필렌 백(polypropylene bag)에 담고 오토클레이브(Autoclave) 안으로 옮겨 121℃에서 60분간 처리하여 살균하고, 40℃에서 건조하여 발효 청미래덩굴 잎을 제조하였다. 차 추출기(Tea Extractor; Damian Tea Co, Anyang, Gyeonggi-Do, Korea)에 열수(약 100℃의 물)를 넣고 여기에 앞에서 제조한 발효 청미래덩굴 잎을 각각 열수 부피 대비 2 중량%(w/v)로 첨가한 후 5분간 우려내어 열수 추출물 제조하였다. 이후, 열수 추출물을 121℃에서 20분간 살균한 후 냉각하고 4℃에서 보관하면서 후술하는 동물실험의 시료로 사용하였다.
3 kg of the mixture of the seedling broth and water was added to 1.5 kg of the above-prepared Cheongmyeonghwa leaf powder, and the mixture was evenly mixed to adjust the total moisture content to about 67%. At this time, the culture solution for fermentation prepared above was used as the seed solution. In addition, the mixed solution of the seed solution and water is prepared by mixing the sterilized water to the seed solution corresponding to 5% (v / w) based on the weight of the seed powder of Cheongmaea vine leaf so that the total amount is 3 liters. Thereafter, a mixture of Cheongmyeonghwa leaf powder, a sterilized liquid and water was placed in a plastic fermentation box (50 cm in length, 70 cm in height, 18 cm in height) at a thickness of about 5 cm, the upper part of the plastic fermentation box was covered with a polyethylene film, Lt; 0 > C for 10 days, 20 days and 30 days, respectively. After fermentation, fermented green tea leaves were placed in a polypropylene bag, transferred into an autoclave, sterilized by treatment at 121 ° C for 60 minutes, and dried at 40 ° C to produce fermented Cyanophyceae. (About 100 ° C water) was put in a tea extractor (Damian Tea Co., Anyang, Gyeonggi-Do, Korea) and the fermented green tea leaves prepared above were added to 2 wt% ), Followed by hot water extraction for 5 minutes. Thereafter, the hot-water extract was sterilized at 121 ° C for 20 minutes, cooled, and stored at 4 ° C to be used as an animal test sample described later.
4. 비발효 청미래덩굴 잎 및 이의 열수 추출물의 제조4. Manufacture of non-fermented Cheongpyeong vine leaf and its hot water extract
앞에서 준비한 청미래덩굴 잎 분말 1.5㎏에 물 3ℓ를 첨가하고 골고루 혼합하여 수분을 평형화시킨 후 이를 폴리프로필렌 백(polypropylene bag)에 담고 오토클레이브(Autoclave) 안으로 옮겨 121℃에서 60분간 처리하여 살균하였다. 이후, 살균된 청미래덩굴 잎 분말을 40℃에서 건조하여 비발효 청미래덩굴 잎을 제조하였다. 차 추출기(Tea Extractor; Damian Tea Co, Anyang, Gyeonggi-Do, Korea)에 열수(약 100℃의 물)를 넣고 여기에 앞에서 제조한 비발효 청미래덩굴 잎을 각각 열수 부피 대비 2 중량%(w/v)로 첨가한 후 5분간 우려내어 열수 추출물을 제조하였다. 이후, 열수 추출물을 121℃에서 20분간 살균한 후 냉각하고 4℃에서 보관하면서 후술하는 동물실험의 시료로 사용하였다.
The water was equilibrated by adding 3 L of water to 1.5 Kg of the above-prepared CheongmyeRa vine leaf powder, and the water was equilibrated. The water was transferred into a polypropylene bag, transferred into an autoclave, and sterilized by treatment at 121 캜 for 60 minutes. Thereafter, the sterilized Cheongmyeongvin leaf powder was dried at 40 DEG C to prepare non-fermented blueberry leaf. (About 100 캜 water) was put into a tea extractor (Damian Tea Co., Anyang, Gyeonggi-Do, Korea), and the non-fermented green tea leaves prepared above were added to 2 wt% v) for 5 minutes and then hot water extract was prepared. Thereafter, the hot-water extract was sterilized at 121 ° C for 20 minutes, cooled, and stored at 4 ° C to be used as an animal test sample described later.
5. 발효 청미래덩굴 잎의 열수 추출물을 이용한 동물실험5. Animal experiment using hot water extract of Fermented Cynomolgus sinensis
(1) 동물실험 방법(1) Animal test method
평균 체중이 20~23g인 4주령의 ICR(Crljori: CD-1), SPF/VAF outbred mice(Orient Ltd., Sungnamsi, Korea)에 기본사료인 5L79 diets(PMI Nutrition, Brentwood, LA)를 급이하여 1주일간 환경에 적응시킨 후, 하기 표 5에서와 같이 4개군(5마리/군)으로 나누어 4주간 사육하였다. 하기 표 5에서 보이는 바와 같이 실험군을 기본사료만을 공급하고 음용수로는 증류수를 급여한 정상대조군(NC), 기본사료만을 급여하고 증류수 대신에 3% 알코올을 대신 급여한 알코올 급여대조군 (AC), 기본사료만을 급여하면서 증류수 대신에 3% 알코올을 함유하는 비발효 청미래덩굴 잎의 열수 추출물을 급여한 군(AC+SC), 기본사료만을 급여하면서 증류수 대신에 3% 알코올을 함유하는 발효 청미래덩굴 잎의 열수 추출물을 급여한 군(AC+FSC)으로 구분하였다.5L79 diets (PMI Nutrition, Brentwood, LA) were fed to four week old ICR (Crljori: CD-1) and SPF / VAF outbred mice (Orient Ltd., Sungnamsi, Korea) with an average weight of 20-23 g After adjusting to the environment for one week, the animals were divided into 4 groups (5 animals / group) as shown in Table 5 below and were bred for 4 weeks. As shown in Table 5, the control group (NC) fed only with the basic diet, the distilled water as the drinking water, the alcohol control group (AC) fed only with the basic diet and the 3% alcohol instead of the distilled water, (AC + SC), which was fed with only 3% alcohol instead of distilled water while feeding only feed, was fed with fermented Cheongpyong vine leaf containing 3% alcohol instead of distilled water (AC + FSC) group.
기본사료는 4℃에서 보관하면서 매일 신선한 식이를 공급하였고, 음용수로 사용한 알코올은 99.98%의 알코올(Sigma Co USA)를 희석하여 사용하였다. 사육장은 stainless steel cage를 사용하였으며, 온도 및 습도는 각각 23℃, 60%로 조정하였다. 명암 주기는 12시간 간격으로 설정하였으며 음용수와 식이는 자유 섭취시켰다.The basic diet was kept at 4 ℃ and fresh diets were supplied daily. Alcohol used as drinking water was diluted with 99.98% alcohol (Sigma Co USA). A stainless steel cage was used for the breeding farm, and temperature and humidity were adjusted to 23 ℃ and 60%, respectively. The light and dark cycle were set at 12 hour intervals.
* SC : 비발효 청미래덩굴 잎의 2% 열수 추출물* SC: 2% hydrothermal extract of non-fermented blue-green vine leaf
* FSC : 발효 청미래덩굴 잎의 2% 열수 추출물
* FSC: 2% hydrothermal extract of fermented Cheongmyeong vine leaf
(2) 측정 항목 및 측정 방법(2) Measurement items and measurement methods
1) 체중, 식이 섭취량, 음용수 섭취량1) Weight, Dietary intake, Drinking water intake
체중, 식이 섭취량 및 음용수 섭취량은 전 실험기간을 통하여 매일 일정한 시간에 측정하였다. 식이효율(feed efficiency ratio, FER)은 같은 기간 동안의 체중 증가량을 동일 기간의 식이 섭취량으로 나눈 값으로 하였다.
Body weight, dietary intake, and drinking water intake were measured at regular intervals throughout the day. The feed efficiency ratio (FER) was defined as the weight gain during the same period divided by the dietary intake for the same period.
2) 분석용 시료의 채취2) Sampling of analytical samples
실험 식이로 4주간 사육한 흰쥐를 물만 주고 24시간 동안 금식시킨 후 에테르(ether) 마취 하에서 복부 대동맥으로부터 채혈한 다음, 빙냉의 생리식염수로 간을 관류하고 장기를 적출 한 후 습기를 제거하여 무게를 측정하였다. 채취한 혈액은 실온에서 응고시킨 다음 4℃, 2,500rpm의 조건으로 15분간 원심분리하여 혈청을 분리한 후 -70℃에서 보관하면서 분석용 시료로 사용하였다.
After rats were fed with water for 4 weeks, they were fasted for 24 hours. After blood was collected from the abdominal aorta under ether anesthesia, the liver was perfused with ice-cold physiological saline and the organs were removed. Respectively. The collected blood was coagulated at room temperature, centrifuged at 4 ° C and 2,500 rpm for 15 minutes, and the serum was separated and stored at -70 ° C for analysis.
3) 혈청 지질(lipid) 함량 측정3) Measurement of serum lipid content
혈청의 중성지질(triglyceride, TG) 함량, 총 콜레스테롤(total cholesterol) 함량 및 HDL-콜레스테롤(HDL-cholesterol) 함량은 kit 시약(AM 157S-K, AM 202-K, AM 203-K, Asanpharm Co., Seoul, Korea)으로 측정하였고, LDL-콜레스테롤(LDL-cholesterol) 함량은 Friedewald 등(1972)의 방법에 준하여 다음과 같은 식으로 계산하였다.The triglyceride (TG) content, total cholesterol content and HDL-cholesterol content of the serum were measured using a kit reagent (AM 157S-K, AM 202-K, AM 203-K, Asanpharm Co.). , Seoul, Korea). LDL-cholesterol content was calculated according to the method of Friedewald et al. (1972) as follows.
LDL-cholesterol = total cholesterol(triglyceride/5+HDL-cholesterol)로 계산하였다.
LDL-cholesterol = total cholesterol (triglyceride / 5 + HDL-cholesterol).
4) 간 조직의 지질(lipid) 함량 측정4) Measurement of lipid content of liver tissue
Folch 등(29)의 방법에 따라 클로로포름(chloroform) : 메탄올(methanol)의 비가 2 : 1인 혼합액에 간 조직의 마쇄액 일정량을 가해 잘 혼합한 다음 방치한 후, 유기용매에 의해 분리된 부분 일정량을 취해 질소가스 존재 하에서 휘발시킨 후 중성지질(triglyceride, TG) 함량은 kit 시약(AM 157S-K, Asanpharm Co., Korea)으로 측정하였고, 총 콜레스테롤(total cholesterol) 함량은 kit 시약(AM 202-K, Asanpharm Co., Korea)으로 측정하였고, 총 지질(total lipid) 함량은 Frings 등(1972)의 방법에 따라 측정하였다.
According to the method of Folch et al. (29), a certain amount of hemolysis of liver tissue was added to a mixture of chloroform: methanol at a ratio of 2: 1, and the mixture was allowed to stand. (AM 157S-K, Asanpharm Co., Korea), and the total cholesterol content of the sample was measured using a kit reagent (AM 202- K, Asanpharm Co., Korea) and the total lipid content was measured according to the method of Frings et al. (1972).
5) 간 조직의 ADH 및 ALDH의 활성측정5) Measurement of activity of ADH and ALDH in liver tissue
알코올 디하이드로게나제(alcohol dehydrogenase: ADH) 및 아세트알데하이드 디하이드로게나제(acetaldehyde dehydrogenase: ALDH)의 활성 측청용 효소원으로 적출한 동물의 간 조직 일정량에 4배량의 빙냉의 0.25M 설탕 용액을 가하여 균질화한 다음 10,000×gf로 20분간 원심분리하여 나온 상징액(postmitochondrial fraction, PMF) 및 미토콘드리아 분획을 사용하였다. 알코올 디하이드로게나제의 활성은 Bernet & Bergermeyer의 방법(Bernet 1983)에 따라 기질인 에탄올과 NAD(nicotinamide adenine dinucleotide)를 첨가하여 반응시키는 동안 생성된 NADH(reduce NAD)의 함량을 340㎚에서 측정하였으며 활성도는 분당 단백 1㎎이 생성시킨 NADH의 양을 nmole로 나타내었다. 아세트알데하이드 디하이드로게나제의 활성은 Lundquist의 방법(Bernet 1983)에 따라 기질인 아세트알데하이드와 NAD를 첨가하여 반응시키는 동안 생성된 NADH의 함량을 파장 340㎚에서 측정하였으며, 활성도는 분당 단백 1㎎이 생성시킨 NADH의 양을 nmole로 나타내었다.
A 4-fold ice-cold 0.25 M sugar solution was added to an amount of liver tissue of an animal extracted with an alcohol source of an alcohol dehydrogenase (ADH) and an acetaldehyde dehydrogenase (ALDH) After homogenization, the postmitochondrial fraction (PMF) and mitochondrial fraction were centrifuged at 10,000 × g f for 20 min. The activity of alcohol dehydrogenase was measured at 340 nm according to the method of Bernet & Bergermeyer (Bernet 1983). The content of NADH (reduce NAD) produced during the reaction of ethanol and NAD (nicotinamide adenine dinucleotide) Activity was expressed in nmole by the amount of NADH produced by 1 mg of protein per minute. The activity of acetaldehyde dehydrogenase was measured at 340 nm in terms of the content of NADH produced during the reaction of acetaldehyde and NAD, which were substrates, according to the method of Lundquist (Bernet 1983). The activity was 1 mg of protein per minute The amount of NADH produced is shown in nmole.
6) 간 조직의 XO(Xanthine oxidase) 및 AO(aldehyde oxidase)의 활성 측정6) Measurement of activity of XO (xanthine oxidase) and AO (aldehyde oxidase) in liver tissues
간 조직으로부터 추출한 PMF를 효소원으로 하여, XO 활성도는 Stirpe와 Della Corte(1969)의 방법에 따라 기질 잔틴(xanthine)을 우릭산(uric acid로 전환하는 정도를 정상대조군에 대한 %로 나타내었으며, AO 활성도는 Rajagopalan 등(1962)의 방법에 따라 기질 NMN (N1-methyl nicotinamide)으로부터 산화된 피리돈(pyridone)을 300 ㎚에서 측정한 다음 정상대조군에 대한 %로 나타내었다.
Extracted from liver tissue The activity of XO was determined by the method of Stirpe and Della Corte (1969), and the degree of conversion of substrate xanthine to uric acid was expressed as% of normal control, and AO activity was measured by Rajagopalan Pyridone oxidized from the substrate NMN (N 1 -methyl nicotinamide) was measured at 300 nm according to the method of Kang et al. (1962) and expressed as% of the normal control group.
7) ALT(alanine aminotransferase) 활성 측정7) Measurement of ALT (alanine aminotransferase) activity
ALT 활성은 kit 시약(Asan Pharm Co., Seoul Korea)으로 측정하였으며 Karmen unit로 나타내었다.
ALT activity was measured by kit reagent (Asan Pharm Co., Seoul Korea) and expressed as Karmen unit.
(3) 측정 결과(3) Measurement result
1) 체중, 식이 섭취량, 음용수 섭취량 등1) Weight, dietary intake, drinking water intake, etc.
하기 표 6은 발효 청미래덩굴 잎의 열수 추출물이 알코올 섭취 실험동물의 체중, 식이 섭취량, 음용수 섭취량 등에 미치는 영향을 나타낸 것이다.Table 6 shows the effect of the hydrothermal extract of fermented Cynomolgus sinensis on the body weight, dietary intake, and drinking water intake of the experimental animals consuming alcohol.
상기 표 6에서 보이는 바와 같이 체중증가량은 알코올을 섭취한 AC, AC+SC, AC+FSC군 모두가 정상군(NC) 보다는 보다 유의적으로 낮았으나 청미래덩굴 잎 열수 추출물을 급여한 군에서는 체중증가량이 알코올만을 섭취한 군에 비해 높아지는 경향을 나타내었으며 이러한 경향은 비발효 청미래덩굴 잎에 비하여 발효 청미래덩굴 잎에서 컸다. 음용수 섭취량은 3% 알코올을 급여한 군들(AC, AC+SC, AC+FSC)에서 증류수를 급여한 NC군에 비하여 높은 경향을 보였으나 발효 청미래덩굴 잎 열수 추출물을 급여한 군은 비발효 청미래덩굴 잎 열수 추출물을 급여한 군 및 알코올만을 급여한 군보다 낮게 나타났다. 이러한 현상은 사육 초기에는 큰 차이를 보이지 않았으나 3% 알코올을 증류수 대신에 급여한 군들에서는 식이 20일 후부터 섭취량의 급격한 증가를 나타내는 것으로 미루어 만성적인 알코올의 식이로 인한 중독현상으로 사료되며, 발효 청미래덩굴 잎 열수 추출물을 급여한 군에서는 그 정도가 크게 완화됨을 알 수 있다.As shown in Table 6, the weight gain was significantly lower in the AC, AC + SC, and AC + FSC groups than in the normal group (NC), but in the group fed with the hydrothermal extract, Were higher than those of the alcohol - only group. The tendency was greater in non - fermented. The intake of drinking water showed a tendency higher than that of NC group fed 3% alcohol (AC, AC + SC, AC + FSC) in distilled water, whereas the group fed fermented Cyanophylla fructus Leaf - water extract group and alcohol - only group. This phenomenon did not show any significant difference at the beginning of breeding, but in the group fed with 3% alcohol instead of distilled water, the intake was increased rapidly from 20 days after the feeding, which is considered to be an addiction due to chronic alcoholic diet. This result shows that the degree of alleviation is greatly reduced in the group fed with the leaf hot water extract.
알코올은 탄수화물이나 단백질에 비하여 칼로리 밀도가 높아 체내 신진대사가 활성화되어 높은 칼로리를 내며, 체내 산소 소비량이 증가하며 호기성 대사의 장애를 유발하고 이로 인한 식욕부진과 영양소 부족현상을 초래하게 되며 체중이 감소하는 현상이 나타난다(Pikaar et al 1897; Gruchow et al 1985; Luciana et al 2009). 만성적인 알코올 섭취는 이러한 현상이 더욱 심하게 나타나며 알코올 대사의 부진으로 알코올 분해물인 아세트알데하이드(acetaldehyde)가 잔류하여 숙취나 소화관 점막의 손상을 유발하며(Cederbaum 1991; Lieber 1997) 식이 섭취량이 감소하는 동시에 체중감소량이 더욱 높아진다.
Alcohol has a higher caloric density than carbohydrates and proteins, so it activates metabolism in the body and increases the calories, increases oxygen consumption in the body, causes aerobic metabolic disorders, causes anorexia and insufficient nutrition, (Pikaar et al 1897; Gruchow et al 1985; Luciana et al 2009). Chronic alcohol consumption is more severe and alcoholic metabolism of acetaldehyde remains, resulting in hangover or gastrointestinal mucosal damage (Cederbaum 1991; Lieber 1997). Dietary intake is decreased while body weight The amount of reduction is further increased.
2) 장기 중량2) Long term weight
하기 표 7은 발효 청미래덩굴 잎의 열수 추출물이 알코올 섭취 실험동물의 장기 중량에 미치는 영향을 나타낸 것이다.Table 7 below shows the effect of the hydrothermal extract of fermented Cynomolgus sinensis L. on the organ weight of an experimental animal consuming alcohol.
상기 표 7에서 보이는 바와 같이 모든 실험군에서 신장, 심장 및 고환의 중량은 차이를 보이지 않았으나 간의 경우는 알코올만 급여한 군(AC)에서 현저한 비대현상을 나타내었다. 그리고 비발효 청미래덩굴 잎 열수 추출물과 발효 청미래덩굴 잎 열수 추출물을 급여한 군은 모두 정상군에는 미치니 못하나 AC군에 비하여는 간의 중량이 뚜렷하게 낮았으며 발효 청미래덩굴 잎 열수 추출물을 급여한 군에서 비발효 청미래덩굴 잎 열수 추출물을 급여한 군에 비하여 더욱 낮은 수치를 나타내었다As shown in Table 7, the weight of the kidney, heart, and testes were not different in all the experimental groups, but the liver showed significant hypertrophy in the group fed only with alcohol (AC). In addition, the weight of liver was significantly lower in the non - fermented group than in the AC group but not in the normal group and in the group fed the fermented panthera vine leaf hydrothermal extract Fermented red ginseng leaves were lower than those of the control group
일반적으로 고지방식이나 고콜레스테롤 식이 또는 만성적으로 음주를 지속하면 체내 영양소대사의 불균형을 초래하게 되며 면역기능이 저하되거나 알코올에 의한 독성 및 이로 인하여 생성된 활성산소종에 의하여 특히 간조직의 손상과 함께 지질함량의 누적으로 간이 비대해 진다(Cederbaum 1991; Lieber CS 1992; Lieber 1997; Mantle과 Preedy 1999; Das 등 2005). 따라서, 상기 표 7에서 나타난 결과는 발효 청미래덩굴 잎 열수 추출물이 만성적인 음주시에도 알코올에 의한 간 조직의 손상을 크게 완화함을 시사한다.
In general, high-fat diet, high cholesterol diet or chronic alcohol consumption can lead to imbalance of nutrient metabolism in the body, resulting in impaired immune function, toxicity due to alcohol, and resulting reactive oxygen species, (Cederbaum 1991; Lieber CS 1992; Lieber 1997; Mantle and Preedy 1999; Das et al. 2005). Therefore, the results shown in the above Table 7 suggest that the hydrothermal extract of Fermented Cyanobacterium japonica significantly alleviates the damage of liver tissue caused by alcohol even during chronic drinking.
3) 혈청 지질 함량3) Serum lipid content
하기 표 8은 발효 청미래덩굴 잎의 열수 추출물이 알코올 섭취 실험동물의 혈청 지질 함량에 미치는 영향을 나타낸 것이다. 하기 표 8에서 각 측정 항목 값은 정상대조군에 대한 %로 나타내었다.Table 8 below shows the effect of the hydrothermal extract of fermented Cynomolgus sinensis on the serum lipid content of the alcohol-consuming experimental animals. The values of each measurement item in Table 8 are shown in% of the normal control group.
상기 표 8에서 보이는 바와 같이 중성지질, 총 콜레스테롤 및 LDL-콜레스테롤 함량은 NC군에 비하여 알코올을 급여한 AC, AC+SC 및 AC+FSC군에서 현저하게 높았다. 그러나 AC+SC 및 AC+FSC군에서는 알코올 섭취로 증가한 지질 성분들의 함량이 뚜렷하게 감소하는 경향을 나타내었으며 그 감소율은 발효 청미래덩굴 잎 열수 추출물을 급여한 군에서 비발효 청미래덩굴 잎 열수 추출물을 급여한 군에 비하여 크게 나타났다. 한편 몸에 이로운 HDL-콜레스테롤 함량은 AC군이 NC군에 비하여 유의적으로 감소하였고 AC+SC군은 AC군과의 유의차를 보이지 않는 반면 AC+FSC군은 AC군과 비교하였을 때 유의적인 증가를 보였다.
As shown in Table 8, the contents of triglyceride, total cholesterol and LDL-cholesterol were significantly higher in the AC, AC + SC and AC + FSC groups fed with alcohol than the NC group. However, in the AC + SC and AC + FSC groups, the content of lipid components increased due to alcohol consumption tended to decrease markedly. The rate of decrease was found to be higher in the group fed with the hydrothermal extract of Cheongpyong Respectively. On the other hand, HDL-cholesterol content, which is beneficial to the body, was significantly decreased in the AC group compared to the NC group, while the AC + SC group showed no significant difference from the AC group, whereas the AC + FSC group showed a significant increase Respectively.
4) 간 조직의 지질 함량4) Lipid content of liver tissue
하기 표 9는 발효 청미래덩굴 잎의 열수 추출물이 알코올 섭취 실험동물의 간 조직 지질 함량에 미치는 영향을 나타낸 것이다. 하기 표 9에서 각 측정 항목 값은 정상대조군에 대한 %로 나타내었다.Table 9 shows the effect of the hydrothermal extract of fermented Cynomolgus sinensis L. on the liver lipid content of the alcohol-fed experimental animals. The values of each measurement item in Table 9 are shown in% of the normal control group.
상기 표 9에서 보이는 바와 같이 총 중성지방 및 총 콜레스테롤의 함량은 청미래덩굴 잎 열수 추출물을 급여한 군에서 알코올만 급여한 군(AC)에 비하여 낮은 함량을 나타내었으며, 특히 발효 청미래덩굴 잎 열수 추출물을 급여한 군에서 비발효 청미래덩굴 잎 열수 추출물을 급여한 군에 비하여 유의적으로 낮았다. 알코올을 만성적으로 섭취하는 경우 알코올 분해효소인 ADH와 ALDH에 의해 NAD가 NADH로 환원되면서 NADH가 과잉으로 축적된다. 축적된 NADH는 지방산의 합성을 촉진시켜 간 조직 내에 지방을 축적하게 하고 지방간이 되게 한다(Cederbaum 1991; Lieber 1991; Lieber 1992; Lieber 1997; Lands 등 1999; Mantle과 Preedy 1999; Das 등 2005). 한편, 간 손상의 지표 효소로 알려진 ALT 활성은 AC군이 NC군에 비하여 현저하게 높았으나 발효 청미래덩굴 잎 열수 추출물 급여군인 AC+FSC군은 AC군에 비하여 ALT 활성이 매우 낮아 간 손상을 예방하거나 지연하는 것으로 나타났다. 이러한 결과로부터 발효 청미래덩굴 잎 열수 추출물이 만성적인 알코올 섭취로 유발할 수 있는 지방간의 발생을 예방하거나 치유하는 효과가 있음을 알 수 있고, 발효 청미래덩굴 잎 열수 추출물의 의약품 소재나 다양한 기능성 식품 소재로의 활용성이 높음을 시사한다.
As shown in Table 9, the total triglyceride and total cholesterol contents were lower in the group fed with the hydrothermal extract of Cheongmaea japonica than in the group fed only with alcohol (AC) And those of the non - fermented bluefin tuna vine leaves were significantly lower than those of the non - fermented vine leaves. When alcohol is chronically ingested, NADH is excessively accumulated as NAD is reduced to NADH by the alcohololytic enzymes ADH and ALDH. The accumulation of NADH accelerates the synthesis of fatty acids, which causes fat accumulation in the liver tissue and leads to fat accumulation (Cederbaum 1991; Lieber 1991; Lieber 1992; Lieber 1997; Lands et al 1999; Mantle and Preedy 1999; The ALT activity, which is known as the indicator enzyme of liver injury, was significantly higher in the AC group than in the NC group. However, the AC + FSC group, which is a fermented group of hydrothermal extracts of fermented vine leaves, Delayed. From these results, it can be seen that the hydrothermal extract of Fermented Chrysanthemum morifolium extract has the effect of preventing or curing the occurrence of fatty liver which can be caused by chronic alcohol intake, and it can be said that the fermented hydrothermal extract of Fermented Chrysanthemum morifolium Suggesting that it is highly applicable.
5) 간 조직에서의 알코올 대사관련 효소 활성5) Enzyme activity related to alcohol metabolism in liver tissue
하기 표 10은 발효 청미래덩굴 잎의 열수 추출물이 알코올 섭취 실험동물의 간 조직에서의 알코올 대사관련 효소 활성에 미치는 영향을 나타낸 것이다. 하기 표 10에서 각 측정 항목 값은 정상대조군에 대한 %로 나타내었다.Table 10 below shows the effect of the hydrothermal extract of fermented Cynomolgus sinensis on alcohol metabolism-related enzymatic activity in liver tissues of alcohol-fed experimental animals. In the following Table 10, the values of each measurement item are expressed as% of the normal control group.
상기 표 10에서 보이는 바와 같이 청미래덩굴 잎 열수 추출물을 급여한 군의 알코올 디하이드로게나제(alcohol dehydrogenase: ADH) 활성 및 아세트알데하이드 디하이드로게나제(acetaldehyde dehydrogenase: ALDH)의 활성은 AC군 및 AC+SC군에 비해 유의적으로 높은 값을 나타내었고, 정상대조군인 NC군의 값과 큰 차이를 보이지 않았다. 일반적으로 체내에 섭취된 에탄올은 ADH에 의해 아세트알데하이드(acetaldehyde)로 전환되고, 아세트알데하이드는 ALDH에 의해 아세트산(acetic acid)으로 산화되며, 궁극적으로 아세틸 조효소(acetyl CoA)로 전환되어 에너지를 생성한다(Lieber 1997). 아세트알데하이드는 에탄올 대사의 중간생성물로 알코올 대사가 원활하지 않는 경우 체내에 잔존하여 조직 손상과 숙취를 유발하며 조직 내 각종 활성산소종을 생성한다(Lieber 1991). 한편, 본 실험에서는 측정하지 않았으나 CYP2E1 효소도 알코올을 아세트알데하이드로 전환시키는 보조적인 역할을 하며 알코올의 만성적 섭취에 의해 간 조직이 손상되면 이 효소의 활성이 높아지는 것으로 알려져 있다(Salmela 등 1998; Albano 2002; Lieber 2008). 상기 표 10에서 나타난 결과로부터 발효 청미래덩굴 잎 열수 추출물은 비발효 청미래덩굴 잎 열수 추출물에 비해 ADH 활성 및 ALDH의 활성을 더 많이 증가시켜 만성적인 음주에서 나타날 수 있는 아세트알데하이드로 인한 숙취와 간 조직 손상을 예방 또는 완화하는데에 큰 효과가 있음을 알 수 있다.
As shown in Table 10, the alcohol dehydrogenase (ADH) activity and the acetaldehyde dehydrogenase (ALDH) activity of the group fed with the hydrothermal extract of Cheongmyeongvin were higher in AC group and AC + SC group compared with the control group, NC group. In general, ethanol in the body is converted to acetaldehyde by ADH, acetaldehyde is converted to acetic acid by ALDH, and ultimately converted to acetyl CoA to generate energy (Lieber 1997). Acetaldehyde is an intermediate product of ethanol metabolism. When alcohol metabolism is not smooth, it remains in the body, causing tissue damage and hangover, and producing various active oxygen species in the tissue (Lieber 1991). Although not measured in this experiment, CYP2E1 enzyme plays an auxiliary role in converting alcohol to acetaldehyde, and it is known that when the liver is damaged by chronic alcohol ingestion, the activity of this enzyme is increased (Salmela et al. 1998; Albano 2002 ; Lieber 2008). From the results shown in the above Table 10, it can be seen that the hydrothermal extract of Fermented Chrysanthemum morifolium increased the activity of ADH activity and ALDH more than that of non-fermented Chrysanthemum morifolium vine leaf hydrolyzate, thereby causing hangover and hepatic tissue damage due to acetaldehyde It can be seen that there is a great effect in preventing or alleviating the disease.
6) 간 조직에서의 XO 활성 및 AO 활성6) XO activity and AO activity in liver tissue
하기 표 11은 발효 청미래덩굴 잎의 열수 추출물이 알코올 섭취 실험동물의 간 조직에서의 XO 활성 및 AO 활성에 미치는 영향을 나타낸 것이다. 하기 표 11에서 각 측정 항목 값은 정상대조군에 대한 %로 나타내었다.Table 11 shows the effect of the hydrothermal extract of fermented Cynomolgus sinensis on the XO activity and AO activity in the liver tissues of the alcohol-consuming experimental animals. The values of each measurement item in Table 11 are shown in% of the normal control group.
상기 표 11에서 보이는 바와 같이 청미래덩굴 잎 열수 추출물을 급여한 군의 XO(Xanthine oxidase) 활성 및 AO(aldehyde oxidase)의 활성은 AC군 및 AC+SC군에 비해 유의적으로 작은 값을 나타내었다. 이러한 결과는 발효 청미래덩굴 잎 열수 추출물이 비발효 청미래덩굴 잎 열수 추출물에 비하여 알코올 급여시 XO 억제 활성 및 AO 억제활성이 큼을 나타내며, 알코올의 만성적 섭취로 인한 체내 활성산소의 생성을 차단하여 알코올로 인한 간 조직 손상을 예방함과 동시에 알코올에 의하여 발생할 수 있는 지방의 축적을 경감시키는 효과가 높음을 시사한다.
As shown in Table 11, the activity of XO (xanthine oxidase) and AO (aldehyde oxidase) in the group fed with the hydrothermal extract of Cheongmyeongvle leaf were significantly lower than those of the AC and AC + SC groups. These results indicate that the hydrothermal extract of Fermented Cranesia japonica has a greater inhibitory activity against XO and AO when fed with alcohol than the non-fermented Cranberry Lycopene hydrothermal extract and inhibits the production of active oxygen due to the chronic intake of alcohol, Suggesting that the effect of alleviating the accumulation of fat, which may be caused by alcohol, is high.
Ⅳ. 제 4차 실험 : 발효 청미래덩굴 잎의 에탄올 추출물, 이의 에틸아세테이트 가용성 IV. Fourth Experiment: Ethanol Extract of Fermented Celery Vine Leaf, Its Ethyl Acetate Solubility
분획물의Fraction
제조 및 주요 기능성 지표항목의 측정 Manufacturing and measurement of key functional metrics items
1. 청미래덩굴 잎 분말의 준비1. Preparation of Chrysanthemum morifolium leaf powder
2012년 10월 대구광역시의 양령 시장에서 구입한 청미래덩굴(Smilax china L.) 잎을 수돗물로 깨끗이 세척한 후 열풍 건조기를 이용하여 40℃에서 충분히 건조시켰다. 이후 건조된 청미래덩굴(Smilax china L.) 잎을 분쇄기(모델명 : HNF-1710; 공급자 : Hanil Electric Co Ltd, Seoul, Korea)를 사용하여 약 40 메쉬(mesh) 정도의 입도 크기로 분쇄한 후 폴리에틸렌 필름으로 밀봉하여 4℃에서 보관하면서 후술하는 발효 시료로 사용하였다.
In October, 2012, the purchase of Smilax ( Smilax china L.) leaves were thoroughly cleaned with tap water and then sufficiently dried at 40 ° C using a hot air dryer. After the drying Smilax China (Smilax china L.) leaf was pulverized to a particle size of about 40 mesh using a pulverizer (model: HNF-1710; supplied by Hanil Electric Co Ltd, Seoul, Korea), sealed with a polyethylene film, And used as a fermentation sample described later.
2. 발효용 균주 배양액의 제조2. Preparation of culture broth for fermentation
발효용 균주로는 이전의 실험에서 청미래덩굴 잎 발효용으로 가장 적합한 것으로 나타난 Aspergillus oryzae KFRI 995 균주를 사용하였다. As a strain for fermentation, the Aspergillus strain oryzae KFRI 995 strain was used.
Aspergillus oryzae KFRI 995 균주를 Bacto™ malt extract(Becton, Dickinson and Company, Sparks, MD, USA)에 이식하고 25℃에서 10일 동안 진탕 배양하였다. 이후, 동일한 배지로 배양액 내 아스퍼질러스 오리재 농도를 108 cells/㎖로 조정하여 Aspergillus oryzae KFRI 995 배양액을 준비하였다.
Aspergillus oryzae KFRI 995 strain was transplanted into Bacto (TM) malt extract (Becton, Dickinson and Company, Sparks, MD, USA) and cultured at 25 ° C for 10 days with shaking. Then, the concentration of aspergillus oryzae in the culture medium was adjusted to 10 8 cells / ml with the same medium, and Aspergillus oryzae KFRI 995 culture medium was prepared.
3. 발효 청미래덩굴 잎, 에탄올 추출물 및 이의 에틸아세테이트 가용성 분획물의 제조3. Preparation of Fermented Cyanophyta Leaf, Ethanol Extract and Its Ethyl Acetate-soluble Fraction
앞에서 준비한 청미래덩굴 잎 분말 1.5㎏에 종균액과 물의 혼합액 3ℓ를 첨가하고 골고루 혼합하여 총 수분 함량을 약 67%가 되게 조정하였다. 이때, 종균액으로는 앞에서 준비한 발효용 균주 배양액을 사용하였다. 또한, 종균액과 물의 혼합액은 청미래덩굴 잎 분말 중량을 기준으로 5%(v/w)에 해당하는 종균액에 살균수를 혼합하여 총 3ℓ가 되도록 조정한 것이다. 이후, 청미래덩굴 잎 분말, 종균액 및 물의 혼합물을 플라스틱 발효 상자(가로 50㎝, 세로 70㎝, 높이 18㎝)에 약 5㎝의 두께로 담고 플라스틱 발효 상자의 상부를 폴리에틸렌 필름으로 덮은 후 약 30℃에서 각각 10일, 20일 및 30일간 발효시켰다. 발효 시간별로 발효가 완료된 청미래덩굴 잎을 폴리프로필렌 백(polypropylene bag)에 담고 오토클레이브(Autoclave) 안으로 옮겨 121℃에서 60분간 처리하여 살균하고, 40℃에서 건조하여 발효 청미래덩굴 잎을 제조하였다. 발효 청미래덩굴 잎 100g을 냉각관이 구비된 추출 장치에 넣고, 여기에 80% 에탄올 수용액 500㎖를 첨가한 후 3시간 동안 가열 추출하였다. 추출액을 여과지(Whatman No 2 filter paper; Whatman International Ltd, Maidstone, England)로 3회 반복해서 여과한 후, 여과액을 40℃에서 감압 농축하고 건조 및 고형화하여 발효 청미래덩굴 잎 분말의 에탄올 추출물을 수득하였다. 이후, 발효 청미래덩굴 잎 분말의 에탄올 추출물에 증류수 500㎖를 가하고 현탁시킨 후, 여기에 에틸아세테이트 500㎖를 가하고 진탕 방치하여 물 가용성 분획층과 에틸아세테이트 가용성 분획층을 분리하였다. 에틸아세테이트 가용성 분획층을 취하고 남은 물 가용성 분획성에 동일한 과정을 2회 반복하여 에틸아세테이트 가용성 분획층을 추가로 획득하였다. 이후, 에틸아세테이트 가용성 분획층을 40℃에서 감압 농축하고 건조 및 고형화하여 발효 청미래덩굴 잎 분말의 에틸아세테이트 가용성 분획물을 수득하였다.
3 kg of the mixture of the seedling broth and water was added to 1.5 kg of the above-prepared Cheongmyeonghwa leaf powder, and the mixture was evenly mixed to adjust the total moisture content to about 67%. At this time, the culture solution for fermentation prepared above was used as the seed solution. In addition, the mixed solution of the seed solution and water is prepared by mixing the sterilized water to the seed solution corresponding to 5% (v / w) based on the weight of the seed powder of Cheongmaea vine leaf so that the total amount is 3 liters. Thereafter, a mixture of Cheongmyeonghwa leaf powder, a sterilized liquid and water was placed in a plastic fermentation box (50 cm in length, 70 cm in height, 18 cm in height) at a thickness of about 5 cm, the upper part of the plastic fermentation box was covered with a polyethylene film, Lt; 0 > C for 10 days, 20 days and 30 days, respectively. After fermentation, fermented green tea leaves were placed in a polypropylene bag, transferred into an autoclave, sterilized by treatment at 121 ° C for 60 minutes, and dried at 40 ° C to produce fermented Cyanophyceae. Fermentation 100 g of Cheongmyeongvle leaf was placed in an extraction apparatus equipped with a cooling tube, 500 ml of an 80% ethanol aqueous solution was added thereto, and the mixture was heated and extracted for 3 hours. The extract was repeatedly filtered three times with Whatman No 2 filter paper (Whatman International Ltd, Maidstone, England), and the filtrate was concentrated under reduced pressure at 40 ° C, dried and solidified to obtain an ethanol extract of the fermented Cyanophyceae leaf powder Respectively. Thereafter, 500 ml of distilled water was added to the ethanol extract of the fermented blueberry leaf powder, suspended in 500 ml of ethyl acetate, and the mixture was shaken and left to separate the water-soluble fraction and the ethyl acetate-soluble fraction. The ethyl acetate soluble fraction layer was further recovered by repeating the same procedure twice as for the water-soluble fractionality remaining after taking the ethyl acetate soluble fraction layer. Thereafter, the ethyl acetate soluble fraction layer was concentrated under reduced pressure at 40 캜, dried and solidified to obtain an ethyl acetate soluble fraction of the fermented Celery seeds.
4. 비발효 청미래덩굴 잎, 에탄올 추출물 및 이의 에틸아세테이트 가용성 분획물의 제조4. Preparation of non-fermented Cheongpyeong vine leaves, ethanol extracts and their ethyl acetate soluble fractions
앞에서 준비한 청미래덩굴 잎 분말 1.5㎏에 물 3ℓ를 첨가하고 골고루 혼합하여 수분을 평형화시킨 후 이를 폴리프로필렌 백(polypropylene bag)에 담고 오토클레이브(Autoclave) 안으로 옮겨 121℃에서 60분간 처리하여 살균하였다. 이후, 살균된 청미래덩굴 잎 분말을 40℃에서 건조하여 비발효 청미래덩굴 잎을 제조하였다. 비발효 청미래덩굴 잎 100g을 냉각관이 구비된 추출 장치에 넣고, 여기에 80% 에탄올 수용액 500㎖를 첨가한 후 3시간 동안 가열 추출하였다. 추출액을 여과지(Whatman No 2 filter paper; Whatman International Ltd, Maidstone, England)로 3회 반복해서 여과한 후, 여과액을 40℃에서 감압 농축하고 건조 및 고형화하여 비발효 청미래덩굴 잎 분말의 에탄올 추출물을 수득하였다. 이후, 비발효 청미래덩굴 잎 분말의 에탄올 추출물에 증류수 500㎖를 가하고 현탁시킨 후, 여기에 에틸아세테이트 500㎖를 가하고 진탕 방치하여 물 가용성 분획층과 에틸아세테이트 가용성 분획층을 분리하였다. 에틸아세테이트 가용성 분획층을 취하고 남은 물 가용성 분획성에 동일한 과정을 2회 반복하여 에틸아세테이트 가용성 분획층을 추가로 획득하였다. 이후, 에틸아세테이트 가용성 분획층을 40℃에서 감압 농축하고 건조 및 고형화하여 비발효 청미래덩굴 잎 분말의 에틸아세테이트 가용성 분획물을 수득하였다.
The water was equilibrated by adding 3 L of water to 1.5 Kg of the above-prepared CheongmyeRa vine leaf powder, and the water was equilibrated. The water was transferred into a polypropylene bag, transferred into an autoclave, and sterilized by treatment at 121 캜 for 60 minutes. Thereafter, the sterilized Cheongmyeongvin leaf powder was dried at 40 DEG C to prepare non-fermented blueberry leaf. 100 g of non-fermented blueberry seedlings were placed in an extraction apparatus equipped with a cooling tube, 500 ml of 80% ethanol aqueous solution was added thereto, and the mixture was heated and extracted for 3 hours. The extract was repeated three times with a filter paper (Whatman International Ltd, Maidstone, England), and the filtrate was concentrated under reduced pressure at 40 ° C, dried and solidified to obtain an ethanol extract of non-fermented blueberry leaf powder . Thereafter, 500 ml of distilled water was added to the ethanol extract of non-fermented blueberry leaf powder, suspended in 500 ml of ethyl acetate, and the mixture was shaken and left to separate the water-soluble fraction and the ethyl acetate-soluble fraction. The ethyl acetate soluble fraction layer was further recovered by repeating the same procedure twice as for the water-soluble fractionality remaining after taking the ethyl acetate soluble fraction layer. Thereafter, the ethyl acetate-soluble fraction layer was concentrated under reduced pressure at 40 캜, dried and solidified to obtain an ethyl acetate-soluble fraction of non-fermented Celery vine leaf powder.
5. 발효 청미래덩굴 잎의 에탄올 추출물 및 이의 에틸아세테이트 가용성 분획물의 주요 기능성 지표항목의 측정5. Determination of major functional index items of ethanol extracts of fermented Celecoxib stem and their soluble fractions of ethyl acetate
(1) 추출 수율(1) extraction yield
도 5는 발효 청미래덩굴 잎 및 비발효 청미래덩굴 잎을 에탄올로 추출하고 에탄올 추출물을 에틸아세테이트로 분획하였을 때의 추출 수율을 나타낸 그래프이다. 일반적으로 플라보노이드를 포함한 폴리페놀성 물질은 상 분리(phase separation)시 주로 에틸아세테트 분획에 모이는 것으로 알려져 있다 (Kim 등 1974). 도 5에서 보이는 바와 같이 청미래덩굴 잎을 아스퍼질러스속 미생물로 발효시킨 후 함수 에탄올로 추출하고 이를 에틸아세테이트로 분획한 결과 비발효 청미래덩굴 잎으로 추출한 경우보다 추출 수율이 높게 나타났다. 이러한 결과는 세포벽과 결합한 상태로 존재하거나 세포질 내에 존재하는 폴리페놀 성분이 발효에 의하여 용출되기 쉬운 상태로 변화되기 때문인 것으로 판단된다. 따라서, 아스퍼질러스속 미생물에 의한 발효는 청미래덩굴 잎에 존재하는 유효성 물질의 이용성을 높일 수 있는 좋은 방법임이 확인되었다.
FIG. 5 is a graph showing the extraction yields of ethanol-extracted and ethanol-extracted fractions of fermented Cyanophyta and non-fermented Cyanoma vine leaves. In general, polyphenolic materials including flavonoids are known to aggregate mainly in ethyl acetate fraction during phase separation (Kim et al., 1974). As shown in FIG. 5, when the leaves of Cheongmyeonghwa were fermented with an aspergillus microorganism, they were extracted with ethanol and fractionated with ethyl acetate. As a result, the extraction yield was higher than that of non-fermented Cheongmyeongvle leaf. These results suggest that the polyphenol component existing in the cell wall or in the cytoplasm is changed into a state that is easily eluted by fermentation. Therefore, it was confirmed that fermentation by aspergillus microorganisms is a good method to improve the availability of the active substance present on the leaf of the Korean blue whale.
(2) 총 플라보노이드 함량 및 총 폴리페놀 함량(2) Total flavonoid content and total polyphenol content
발효 청미래덩굴 잎의 에탄올 추출물 및 이의 에틸아세테이트 가용성 분획물의 총 플라보노이드 함량 및 총 폴리페놀 함량을 제 1차 실험의 분석방법과 동일한 방법으로 측정하였다.The total flavonoid content and total polyphenol content of the ethanol extracts of the fermented Cheongpyong vine leaves and the ethyl acetate soluble fraction thereof were measured by the same method as the analysis method of the first experiment.
도 6은 발효 청미래덩굴 잎 및 비발효 청미래덩굴 잎의 에탄올 추출물 및 이의 에틸아세테이트 가용성 분획물에 존재하는 총 폴리페놀 함량을 나타낸 그래프이고, 도 7은 발효 청미래덩굴 잎 및 비발효 청미래덩굴 잎의 에틸아세테이트 가용성 분획물에 존재하는 총 플라보노이드 함량 : 총 폴리페놀 함량의 비를 백분율로 나타낸 것이다. 도 6 내지 도 7에서 보이는 바와 같이 발효 청미래덩굴 잎의 에탄올 추출물은 비발효 청미래덩굴 잎의 에탄올 추출물에 비해 총 폴리페놀 함량이 낮았으나, 발효 청미래덩굴 잎의 에틸아세테이트 가용성 분획물은 비발효 청미래덩굴 잎의 에틸아세테이트 가용성 분획물에 비해 총 폴리페놀 함량이 높았고, 총 플라보노이드 대 총 폴리페놀의 비율은 약 2.88배 정도 증가하였다. 이와 같은 결과는 아스퍼질러스속 미생물에 의한 청미래덩굴 잎의 발효에 의해 청미래덩굴 잎에 존재하는 폴리페놀 성분의 일부가 플라보노이드로 전환됨과 동시에 글리코사이드(glycoside) 형태의 플라보노이드에서 당류 부분이 분해 또는 제거되었기 때문인 것으로 사료되며 그로 인해 발효 청미래덩굴 잎의 기능성이 향상된 것으로 판단된다(Hamzeh-Mivehroud 2013).
FIG. 6 is a graph showing the total polyphenol contents in the ethanol extracts of fermented Cheongpyong vine leaves and non-fermented Cheongmyeon vine leaves and their ethyl acetate soluble fractions, and FIG. 7 is a graph showing the total polyphenol contents in fermented Cheongpyong vine leaves and ethyl acetate The percentage of total flavonoid content: total polyphenol content present in the soluble fraction is expressed as a percentage. As shown in FIGS. 6 to 7, the total ethanol content of the fermented Cheongpyong vine leaf was lower than that of the non-fermented Cheongpyong vine leaf, but the ethyl acetate soluble fraction of the fermented Cheongpyong vine leaf was higher than that of non- , And the ratio of total flavonoids to total polyphenols was increased about 2.88 times compared to that of ethyl acetate soluble fraction of ethyl acetate. These results indicate that some of the polyphenols present in the leaves of Cheongmyeongjae were converted into flavonoids by the fermentation of Aspergillus microorganisms and that the saccharide moieties were decomposed or removed from the glycoside type flavonoids (Hamzeh-Mivehroud 2013). It is believed that the function of the fermented Cynomolgus sinensis leaf is improved.
(3) XOIA(Xanthine oxidase inhibitory activity) 및 AOIA(aldehyde oxidase inhibitory activity)(3) Xanthine oxidase inhibitory activity (XOIA) and aldehyde oxidase inhibitory activity (AOIA)
발효 청미래덩굴 잎의 에틸아세테이트 가용성 분획물의 XO 억제 활성 및 AO 억제 활성을 제 1차 실험의 분석방법과 동일한 방법으로 측정하였다.The XO inhibitory activity and the AO inhibitory activity of the ethyl acetate soluble fraction of fermented Cynomolgus sinensis leaf were measured by the same method as in the first experiment.
하기 표 12는 발효 청미래덩굴 잎의 에틸아세테이트 가용성 분획물의 XO 억제 활성 및 AO 억제 활성을 각각 비발효 청미래덩굴 잎의 에틸아세테이트 가용성 분획물의 XO 억제 활성 및 AO 억제 활성에 대한 백분율로 나타낸 것이다.Table 12 shows the XO inhibitory activity and the AO inhibitory activity of the ethyl acetate-soluble fraction of the fermented Cynomolgus sinensis leaf, respectively, as a percentage of the XO inhibitory activity and the AO inhibitory activity of the ethyl acetate-soluble fraction of non-fermented Cynomolium cv.
상기 표 12에서 보이는 바와 같이 발효 청미래덩굴 잎의 에틸아세테이트 가용성 분획물은 비발효 청미래덩굴 잎의 에틸아세테이트 가용성 분획물에 비해 XO 억제 활성은 약 1.90배, AO 억제 활성은 약 2.37배 정도 높은 것으로 나타났다. 상기 결과로부터 발효 청미래덩굴 잎 또는 이의 추출물은 활성산소 생성계 효소인 XO와 AO를 효과적으로 억제하여 요산의 과다 생성 방지에 의한 요통의 예방 내지 완화 기능, 더 나아가 알코올 및 고지방 섭취에 의한 간 손상의 예방 내지 치유 기능이 비발효 청미래덩굴 잎 또는 이의 추출물에 비하여 현저하게 증진됨을 알 수 있다.
As shown in Table 12, the ethyl acetate-soluble fraction of the fermented Cheongpyong vine leaves showed an XO inhibitory activity of about 1.90 times and an AO inhibitory activity of about 2.37 times higher than the ethyl acetate-soluble fraction of non-fermented Cynomolium cv. From the above results, it was found that the fermented blueberry leaf or its extract effectively inhibited XO and AO, which are active oxygen generating enzymes, to prevent or alleviate back pain by preventing overproduction of uric acid, and further to prevent liver damage by alcohol and high fat intake And the healing function is remarkably enhanced as compared with the non-fermented Cynomolgus sinensis or its extract.
Ⅴ. 제 5차 실험 : 발효 청미래덩굴 잎의 에틸아세테이트 가용성 Ⅴ. Fifth experiment: Ethyl acetate solubility of fermented blue-green vine leaves
분획물의Fraction
간 손상 개선 효과 평가 Evaluation of liver damage improvement effect
1. 동물실험 방법1. Animal test method
평균 체중이 20~23g인 4주령의 ICR(Crljori: CD-1), SPF/VAF outbred mice(Orient Ltd., Sungnamsi, Korea)에 기본사료인 5L79 diets(PMI Nutrition, Brentwood, LA)를 급이하여 1주일간 환경에 적응시킨 후, 4개군(5마리/군)으로 나누어 4주간 사육하였다. 구체적으로 실험군을 올리브오일로 처리한 후 기본사료를 급여한 정상대조군(NC), 사염화탄소로 처리하여 간 손상을 유도한 후 기본사료를 급여한 군(CB), 사염화탄소로 처리하여 간 손상을 유도한 후 제 4차 실험에서 수득한 비발효 청미래덩굴 잎의 에틸아세테이트 가용성 분획물을 기본사료에 0.5% 혼합하여 급여한 군(CNS) 및 사염화탄소로 처리하여 간 손상을 유도한 후 제 4차 실험에서 수득한 발효 청미래덩굴 잎의 에틸아세테이트 가용성 분획물을 기본사료에 0.5% 혼합하여 급여한 군(CFS)으로 구분하여 실험하였다. 사육장은 stainless steel cage를 사용하였으며, 온도 및 습도는 각각 20±2℃, 60±5%로 조정하였다. 명암 주기는 12시간 간격으로 설정하였으며 음용수와 식이는 자유 섭취시켰다.5L79 diets (PMI Nutrition, Brentwood, LA) were fed to four week old ICR (Crljori: CD-1) and SPF / VAF outbred mice (Orient Ltd., Sungnamsi, Korea) with an average weight of 20-23 g After adjusting to the environment for 1 week, the animals were divided into 4 groups (5 animals / group) for 4 weeks. Specifically, the experimental group was treated with olive oil, treated with normal control (NC), and treated with carbon tetrachloride to induce liver damage, and treated with carbon tetrachloride (CB) (CNS) and carbon tetrachloride by mixing 0.5% of the ethyl acetate-soluble fraction of the non-fermented Cynomolgus sinensis leaf obtained in the fourth experiment with 0.5% of the basic feed and inducing liver damage, The ethyl acetate soluble fraction of fermented Cynomolgus sinensis leaf was mixed with 0.5% of the basic feed and divided into CFS group. A stainless steel cage was used for the breeding farm, and the temperature and humidity were adjusted to 20 ± 2 ℃ and 60 ± 5%, respectively. The light and dark cycle were set at 12 hour intervals.
CB군, CNS군 및 CFS군의 경우 50% 사염화탄소와 올리브오일의 1:1로 혼합물을 실험 개시일부터 쥐 무게 100g 당 0.1㎖의 용량으로 매일 1회씩 2일 동안 복강에 주사하여 간 손상을 유도하였다. 반면, NS군은 올리브오일만 복강에 주사하였다.
For the CB, CNS, and CFS groups, a 1: 1 mixture of 50% carbon tetrachloride and olive oil was injected intraperitoneally for 1 day into the abdominal cavity at a dose of 0.1 ml per 100 g of rat weight starting from the day of the experiment to induce liver injury . On the other hand, the NS group injected only olive oil into the abdominal cavity.
2. 측정 항목 및 측정 방법2. Measurement items and measurement methods
(1) 체중, 식이 섭취량, 식이효율(1) body weight, dietary intake, dietary efficiency
체중, 식이 섭취량 및 음용수 섭취량은 전 실험기간을 통하여 매일 일정한 시간에 측정하였다. 식이효율(feed efficiency ratio, FER)은 같은 기간 동안의 체중 증가량을 동일 기간의 식이 섭취량으로 나눈 값으로 하였다.
Body weight, dietary intake, and drinking water intake were measured at regular intervals throughout the day. The feed efficiency ratio (FER) was defined as the weight gain during the same period divided by the dietary intake for the same period.
(2) 분석용 시료의 채취(2) Sampling of analytical samples
실험 식이로 4주간 사육한 흰쥐를 물만 주고 24시간 동안 금식시킨 후 에테르(ether) 마취 하에서 복부 대동맥으로부터 채혈한 다음, 빙냉의 생리식염수로 관류하고 간 조직을 적출하였다. 적출한 간 조직 일정량에 4배량의 빙냉의 0.25M 설탕 용액을 가하고 마쇄하여 균질화한 다음 10,000×gf로 20분간 원심분리하여 나온 상징액(postmitochondrial fraction, PMF)을 취해 효소 활성도 측정에 이용하였다.
Rats fed the diet for 4 weeks were fasted for 24 hours with water only. Blood was collected from the abdominal aorta under ether anesthesia, and then perfused with ice - cold physiological saline and liver tissues were extracted. A 4-fold ice-cold 0.25 M sugar solution was added to the extracted liver tissue, homogenized, and the postmitochondrial fraction (PMF) obtained by centrifugation at 10,000 × g f for 20 minutes was used for enzyme activity measurement.
(3) ALT 활성 측정(3) Measurement of ALT activity
ALT 활성은 kit 시약(Asan Pharm Co., Seoul Korea)으로 측정하였으며 Karmen unit로 나타내었다.
ALT activity was measured by kit reagent (Asan Pharm Co., Seoul Korea) and expressed as Karmen unit.
(4) XO(Xanthine oxidase) 활성 및 AO(aldehyde oxidase)의 활성 측정(4) Measurement of activity of XO (xanthine oxidase) and AO (aldehyde oxidase)
간 조직으로부터 추출한 PMF를 효소원으로 하여, XO 활성도는 Stirpe와 Della Corte(1969)의 방법에 따라 기질 잔틴(xanthine)을 우릭산(uric acid로 전환하는 정도를 정상대조군에 대한 %로 나타내었으며, AO 활성도는 Rajagopalan 등(1962)의 방법에 따라 기질 NMN (N1-methyl nicotinamide)으로부터 산화된 피리돈(pyridone)을 300 ㎚에서 측정한 다음 정상대조군에 대한 %로 나타내었다.
Extracted from liver tissue The activity of XO was determined by the method of Stirpe and Della Corte (1969), and the degree of conversion of substrate xanthine to uric acid was expressed as% of normal control, and AO activity was measured by Rajagopalan Pyridone oxidized from the substrate NMN (N 1 -methyl nicotinamide) was measured at 300 nm according to the method of Kang et al. (1962) and expressed as% of the normal control group.
(5) 글루타치온(glutathione, GSH) 함량 및 과산화지질(lipid peroxide, LPO) 함량 측정(5) Determination of glutathione (GSH) content and lipid peroxide (LPO) content
간 조직의 GSH 함량 및 LPO의 함량은 Ellman(1959) 및 Satho(14)의 방법에 따라 각각 측정하였다. 단백질의 함량은 Lowry(1951)법에 따라 표준품으로 bovine serum albumin을 사용하여 측정하였다.
GSH and LPO contents of liver tissues were measured by Ellman (1959) and Satho (14), respectively. Protein content was measured using bovine serum albumin as a standard product according to Lowry (1951) method.
3. 측정 결과3. Measurement results
하기 표 13은 발효 청미래덩굴 잎의 에틸아세테이트 가용성 분획물이 간 손상이 유도된 실험동물의 체중, 식이 섭취량, ALT 활성, XO 활성 등에 미치는 영향을 나타낸 것이다.Table 13 shows the effect of the ethyl acetate-soluble fraction of fermented blueberry leaf on the body weight, dietary intake, ALT activity, and XO activity of liver injury-induced experimental animals.
1) 체중, 식이 섭취량 및 식이효율1) Weight, dietary intake and diet efficiency
NC군의 체중 증가량은 주당 1.74g을 나타내었으나 CB군과 CNS군은 체중이 감소하는 결과를 나타내었으며, CFS군은 NC군에는 미치지 않으나 체중이 증가하는 경향을 보였다. 식이 섭취량은 CB군이 NC군에 비하여 다소 높았으나 사염화탄소 처리군 간의 차이는 없었다. CB군과 CNS군은 사염화탄소의 독성에 의하여 간 조직이 손상되고 그로 인해 체중이 감소한 것으로 보이나 CFS군은 CNS군에 비하여 사염화탄소에 대한 해독 효과가 커서 체중이 감소하지 않은 것으로 판단된다.
The weight gain of the NC group was 1.74 g per week, but the CB group and the CNS group showed a decrease in body weight. The CFS group showed a tendency to increase in body weight, though not in the NC group. Dietary intakes were slightly higher in the CB group than in the NC group, but there was no difference between the groups in the carbon tetrachloride treatment group. CB group and CNS group showed liver damage due to toxicity of carbon tetrachloride and decreased body weight. However, CFS group did not decrease weight because of detoxification effect on carbon tetrachloride compared with CNS group.
2) 간 중량과 ALT 활성2) liver weight and ALT activity
간의 중량은 CB군이 NC군에 비하여 59.52%가 증가하였으나, CNC군은 CB군에 비하여 23.89%가 감소하였고, CFS군은 CB군에 비하여 28.36%가 감소하였으며, 발효 청미래덩굴 잎의 에틸아세테이트 가용성 분획물을 급여한 군에서 간의 중량이 NC군과 비슷한 값을 나타내었다. ALT 활성에서도 CB군은 NC군에 비하여 2.90배가 높았으나 CNS군과 CFS군은 CB군에 비하여 각각 17.76% 및 28.66% 정도가 유의적으로 감소하여, 발효 청미래덩굴 잎의 에틸아세테이트 가용성 분획물을 급여한 군에서 상대적으로 낮은 ALT 활성을 보였다. 간이 손상되면 ALT의 활성이 높아지며, ALT의 활성도는 간 손상의 지표로 활용되고 있다. 이러한 결과로부터 발효 청미래덩굴 잎의 에틸아세테이트 가용성 분획물은 사염화탄소와 같은 물질들의 간 독성을 해독 효과가 매우 크고 사염화탄소 등에 의한 간 손상을 효과적으로 예방하거나 치료할 수 있음을 알 수 있다.
The weight of the liver was increased by 59.52% in the CB group compared to the NC group, but decreased by 23.89% in the CNC group and 28.36% in the CFS group compared with the CB group. Ethyl acetate solubility The liver weights were similar to those of NC group. ALT activity was higher in the CB group than in the NC group, but the CNS group and the CFS group were significantly decreased by 17.76% and 28.66%, respectively, compared to the CB group, and the ethyl acetate soluble fraction of the fermented Cynomolgus japonica leaf was fed ALT activity was relatively low in the control group. Liver damage increases the activity of ALT, and ALT activity is used as an index of liver damage. From these results, it can be seen that the ethyl acetate-soluble fraction of the fermented Cynthia rugosa leaf is very effective in detoxifying hepatotoxicity of substances such as carbon tetrachloride and effectively preventing or treating liver damage caused by carbon tetrachloride and the like.
3) 간 조직의 GSH 함량 및 LPO 함량과 XO 활성 및 AO 활성3) GSH content, LPO content, XO activity and AO activity in liver tissues
GSH 함량은 사염화탄소를 투여한 모든 실험군에서 뚜렷한 변화가 없었다. 과산화지질(LPO)의 함량은 사염화탄소를 투여한 모든 실험군에서 전반적으로 NC군에 비하여 높은 함량을 나타내었으나, CFS군에서 유의적으로 낮았다. 또한, XO 활성 및 AO의 활성은 CB군 및 CNS군에서 NC군보다 높았으나 CFS군은 NC군과 비슷한 값을 나타내어 유의적으로 낮았다. 사염화탄소와 같은 약물에 의하여 간이 손상되면 활성산소계 효소인 XO와 AO의 활성이 높아지며(Nishino & Tamura 1991), 이 결과 생성된 활성산소종(ROS)은 체내 지질을 산화시켜 지질과산화물(LPO)을 생성한다. 일반적으로 LPO의 생성이 많아지면 ROS 소거계로 작용하는 GSH와 같은 물질이 작용하게 된다. 이상의 결과로부터 발효 청미래덩굴 잎의 에틸아세테이트 가용성 분획물은 비발효 청미래덩굴 잎의 에틸아세테이트 가용성 분획물보다 XO 활성 및 AO 활성을 크게 억제하여 사염화탄소와 같은 간 손상을 일으키는 독성물질의 해독효과가 높고, 간 손상을 감소시키는 효과가 큼을 알 수 있다.
GSH content was not significantly changed in all experimental groups treated with carbon tetrachloride. The lipid peroxidation (LPO) content was higher in all experimental groups treated with carbon tetrachloride than in the NC group, but significantly lower in the CFS group. The activities of XO and AO were higher in the CB and CNS groups than in the NC group, but significantly lower in the CFS group than in the NC group. When liver damage is mediated by drugs such as carbon tetrachloride, the activity of active oxygen species XO and AO increases (Nishino & Tamura 1991), and the resulting reactive oxygen species (ROS) oxidize lipids in the body to produce lipid peroxides do. Generally, when the production of LPO is increased, a substance such as GSH which acts as an ROS scavenging agent acts. From the above results, the ethyl acetate-soluble fraction of fermented Cheongpyong vine leaves significantly inhibited XO activity and AO activity than the ethyl acetate-soluble fraction of non-fermented Cheongpyong vine leaves, so that toxic substances causing liver damage such as carbon tetrachloride were highly detoxified, It can be seen that the effect of decreasing the amount of water is great.
Ⅵ. 제 6차 실험 : 발효 청미래덩굴 잎을 이용한 발효 차 및 기능성 음료의 제조VI. Sixth Experiment: Production of Fermented Tea and Functional Beverage Using Fermented Celadon Vine Leaves
1. 발효 차의 제조 및 관능 검사
1. Manufacturing and Sensory Evaluation of Fermented Tea
(1) 발효 차의 제조(1) Production of fermented tea
제조예 1.Production Example 1
제 2차 실험에서 Aspergillus oryzae KFRI 995 균주로 10일간 발효시켜 제조한 발효 청미래덩굴 잎을 200℃의 덖음솥에서 약 40초간 덖은 후 2g씩 티백에 담아 발효 차를 제조하였다.
In the second experiment, Aspergillus oryzae KFRI 995 fermented for 10 days. The fermented green tea leaves were fermented for about 40 seconds in a 200 ° C pot.
제조예 2.Production Example 2
제 2차 실험에서 Aspergillus oryzae KFRI 995 균주로 10일간 발효시켜 제조한 발효 청미래덩굴 잎을 200℃의 덖음솥에서 약 40초간 덖은 후 1g씩 티백에 담았다. 옥수수 줄기 분말 및 꾸지뽕 분말을 각각 떡음 처리한 후 1:1의 중량비로 혼합하고, 발효 청미래덩굴 잎이 담긴 티백에 이를 1g씩 담아 발효 차를 제조하였다.
In the second experiment, Aspergillus oryzae KFRI 995 fermented by fermentation for 10 days. The fermented Cheongmyeonghwa leaf was melted for about 40 seconds in a 200 ℃ oven and then put in 1 g of tea bag. Cornstarch powder, and coconut powder were mixed with each other at a weight ratio of 1: 1, and 1 g of each of them was added to a tea bag containing fermented blueberry leaf vine leaves to prepare a fermented tea.
(2) 발효 차의 관능검사(2) Sensory evaluation of fermented tea
제조예 1 및 제조예 2에서 제조한 발효 차를 95~100℃의 열수로 약 3분간 침출하여 관능검사용 시료로 사용하였다. 침출 차에 대해 훈련된 관능 요원 25명을 대상으로 종합적인 기호도(overall acceptability)를 5점 측도법으로 평가하였다(Herbert & Jeol 1993). 5점 측도법의 평가 점수 기준은 아주 나쁘다 : 1점; 나쁘다 : 2점; 보통이다 : 3점; 좋다 : 4점; 아주 좋다 : 5점으로 하였다. 관능검사 결과 제조예 1에서 제조한 발효 차의 종합적인 기호도는 3.54인 것으로 나타났고, 제조예 2에서 제조한 발효 차의 종합적인 기호도는 3.91인 것으로 나타났다.
The fermented tea prepared in Preparation Examples 1 and 2 was leached with hot water at 95 to 100 ° C for about 3 minutes and used as a sample for sensory evaluation. Overall acceptability was assessed by the five point scoring method for 25 sensory agents trained in leach tea (Herbert & Jeol 1993). The five point scoring method scoring standard is very bad: 1 point; Bad: 2 points; Usually: 3 points; Good: 4 points; Very good: 5 points. As a result of sensory evaluation, the overall acceptability of the fermented tea prepared in Preparation Example 1 was found to be 3.54, and the overall acceptability of the fermented tea prepared in Preparation Example 2 was found to be 3.91.
2. 기능성 음료의 제조 및 관능 검사
2. Manufacturing and Sensory Evaluation of Functional Beverages
(1) 기능성 음료의 제조(1) Production of functional beverage
제조예 3.Production Example 3
제 2차 실험에서 Aspergillus oryzae KFRI 995 균주로 10일간 발효시켜 제조한 발효 청미래덩굴 잎을 200℃의 덖음솥에서 약 40초간 덖은 후 여기에 물을 가하여 발효 청미래덩굴 잎의 농도가 1%(w/v)인 현탁액을 제조하였다. 이후, 발효 청미래덩굴 잎 현탁액을 약 90~100℃로 가열하고 약 10분 동안 추출하였다. 추출액을 살균 면포로 여과하여 기능성 음료를 제조하였다.
In the second experiment, Aspergillus oryzae KFRI 995 fermented for 10 days in a fermentation pan at a temperature of 200 ° C for about 40 seconds and then water was added thereto to prepare a suspension having a concentration of 1% (w / v) . Thereafter, the suspension of the fermented blueberry leaf vine leaves was heated to about 90 to 100 DEG C and extracted for about 10 minutes. The extract was filtered with a pasteurized cotton to prepare a functional beverage.
제조예 4.Production Example 4
제 2차 실험에서 Aspergillus oryzae KFRI 995 균주로 10일간 발효시켜 제조한 발효 청미래덩굴 잎을 200℃의 덖음솥에서 약 40초간 덖은 후 여기에 물을 가하여 발효 청미래덩굴 잎의 농도가 1%(w/v)인 현탁액을 제조하였다. 이후, 발효 청미래덩굴 잎 현탁액을 약 90~100℃로 가열하고 약 5~10분간 유지하여 추출하였다. 추출액을 살균 면포로 여과하여 베이스 원액을 제조하였다. 또한, 옥수수 뿌리, 흑삼에 각각 약 90~100℃의 열수를 첨가하여 2%(w/v) 농도로 조정하고 약 2시간 동안 추출하였다. 이후, 베이스 원액 91㎖, 옥수수 뿌리 추출액 4㎖, 흑삼 추출액 4㎖, 사양벌꿀 3g, 비타민 혼합액(비타민 B1 1g, 비타민 B12 1g, 니코틴아마이드 1g, 비타민 C 1g, 이노시톨 1g 및 타우린 1g을 증류수 99㎖에 녹인 용액) 1㎖을 혼합하여 기능성 음료를 제조하였다.
In the second experiment, Aspergillus oryzae KFRI 995 fermented for 10 days in a fermentation pan at a temperature of 200 ° C for about 40 seconds and then water was added thereto to prepare a suspension having a concentration of 1% (w / v) . Thereafter, the suspension of the fermented Cynomolgus sinensis leaf was heated to about 90 to 100 캜 and maintained for about 5 to 10 minutes. The extract solution was filtered with a sterilized cotton cloth to prepare a base stock solution. In addition, corn roots and black ginseng were adjusted to a concentration of 2% (w / v) by adding hot water of about 90 to 100 ° C, respectively, and extracted for about 2 hours. Thereafter, 91 ml of base stock solution, 4 ml of corn root extract, 4 ml of black ginseng extract, 4 ml of specimen honey, 3 g of vitamins mixture (1 g of vitamin B1, 1 g of vitamin B12, 1 g of nicotinamide, 1 g of vitamin C, 1 g of inositol and 1 g of taurine, ) Were mixed to prepare a functional beverage.
(2) 기능성 음료의 관능검사(2) Sensory evaluation of functional beverages
제조예 3 및 제조예 4에서 제조한 기능성 음료에 대해 훈련된 관능 요원 25명을 대상으로 종합적인 기호도(overall acceptability)를 5점 측도법으로 평가하였다(Herbert & Jeol 1993). 5점 측도법의 평가 점수 기준은 아주 나쁘다 : 1점; 나쁘다 : 2점; 보통이다 : 3점; 좋다 : 4점; 아주 좋다 : 5점으로 하였다. 관능검사 결과 제조예 3에서 제조한 기능성 음료의 종합적인 기호도는 3.52인 것으로 나타났고, 제조예 4에서 제조한 기능성 음료의 종합적인 기호도는 4.07인 것으로 나타났다.
The overall acceptability was evaluated by a five point scoring method (Herbert & Jeol, 1993) in 25 sensory agents trained in the functional beverages prepared in Preparation Examples 3 and 4. The five point scoring method scoring standard is very bad: 1 point; Bad: 2 points; Usually: 3 points; Good: 4 points; Very good: 5 points. As a result of sensory evaluation, the overall acceptability of the functional beverage prepared in Preparation Example 3 was found to be 3.52, and that of the functional beverage prepared in Preparation Example 4 was 4.07.
이상에서와 같이 본 발명을 상기의 실시예를 통해 설명하였지만 본 발명이 반드시 여기에만 한정되는 것은 아니며 본 발명의 범주와 사상을 벗어나지 않는 범위 내에서 다양한 변형실시가 가능함은 물론이다. 따라서, 본 발명의 보호범위는 본 발명에 첨부된 특허청구의 범위에 속하는 모든 실시 태양을 포함하는 것으로 해석되어야 한다.
While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims. Accordingly, the scope of protection of the present invention should be construed as including all embodiments falling within the scope of the appended claims.
[참고 문헌][references]
1. Albano E. 2002. Free-radicals and alcohol-induced liver injury. In: Sherman, CDIN, Preedy VR, Walson RR Eds., Ethanol and Liver. Taylor and Francis, London, pp. 153190.1. Albano E. 2002. Free-radicals and alcohol-induced liver injury. In: Sherman, CDIN, Preedy VR, Walson RR Eds., Ethanol and Liver. Taylor and Francis, London, pp. 153190.
2. Ali S, Pawa S, Naime M, Prasad R, Ahmad T, Farooqui H, Zafar H. 2008. Role of mammalian cytosolic molybdenum Fe-S flavin hydroxylases in hepatic injury. Life Sci 82: 780-788.2. Ali S, Pawa S, Naime M, Prasad R, Ahmad T, Farooqui H, Zafar H. 2008. Role of mammalian cytosolic molybdenum Fe-S flavin hydroxylases in hepatic injury. Life Sci 82: 780-788.
3. Al-Salmy HS. 2001. Individual variation in hepatic aldehyde oxidase activity. IUBMB Life 51(4): 249-253.3. Al-Salmy HS. 2001. Individual variation in hepatic aldehyde oxidase activity. IUBMB Life 51 (4): 249-253.
Angayarkanni J, Palaniswamy M, Murugesan S, Swaminathan K. 2002. Improvement of tea leaves fermentation with Aspergillus spp. pectinase. J Biosci Bioeng 94(4): 299-303.Angayarkanni J, Palaniswamy M, Murugesan S, Swaminathan K. 2002. Improvement of tea leaves fermentation with Aspergillus spp. pectinase. J Biosci Bioeng 94 (4): 299-303.
4. Anzai N, Enomoto A, Endou H. 2005. Renal urate handling: clinical relevance of recent advances. Current Rheumatology Reports 7: 22734.4. Anzai N, Enomoto A, Endou H. 2005. Renal urate handling: clinical relevance of recent advances. Current Rheumatology Reports 7: 22734.
5. Arakawa H, Maeda M, Okubo S, Shimamura T. 2004. Role of hydrogen peroxide in bactericidal action of catechin. Biol Pharm Bull 27: 277-281. 5. Arakawa H, Maeda M, Okubo S, Shimamura T. 2004. Role of hydrogen peroxide in bactericidal action of catechin. Biol Pharm Bull 27: 277-281.
6. Banerjee A, Dasgupta N, De B. 2005. In vitro study of antioxidant activity of Syzygium cumini fruit. Food Chem 90: 727-733. 6. Banerjee A, Dasgupta N, De B. 2005. In vitro study of antioxidant activity of Syzygium cumini fruit. Food Chem 90: 727-733.
7. Beedham C. 1987. Molybdenum hydroxylases: biological distribution and substrate- inhibitor specificity. Prog Med Chem 24: 85-121.7. Beedham C. 1987. Molybdenum hydroxylases: biological distribution and substrate-inhibitor specificity. Prog Med Chem 24: 85-121.
8. Benzie IF, Strain JJ. 1996. The ferric reducing ability of plasma (FRAP) as a measure of antioxidant power: the FRAP assay. Anal Biochem 239: 7076. 8. Benzie IF, Strain JJ. 1996. The ferric reducing ability of plasma (FRAP) as a measure of antioxidant power: the FRAP assay. Anal Biochem 239: 7076.
9. Bernet E. 1983. Methods of enzymatic analysis. Vch Pub; 3rd edition pp. 1506-1509.9. Bernet E. 1983. Methods of enzymatic analysis. Vch Pub; 3rd edition pp. 1506-1509.
10. Blois MS. 1958. Antioxidant determination by the use of a stable free radical. Nature 181: 1199-1200.10. Blois MS. 1958. Antioxidant determination by the use of a stable free radical. Nature 181: 1199-1200.
11. Carr MC (2003) The emergence of the metabolic syndrome with menopause. J Clin Endocrinol Metab 88(6): 2404-2411.11. Carr MC (2003) The emergence of the metabolic syndrome with menopause. J Clin Endocrinol Metab 88 (6): 2404-2411.
12. Caulfield MJ, Munroe PB, OD, Witkowska K, Charchar FJ, Doblado M, Evans S, Eyheramendy S, Onipinla A, Howard P, Shaw-Hawkins S, Dobson RJ, Wallace C, Newhouse SJ, Brown M, Connell JM, Dominiczak A, Farrall M, Lathrop GM, Samani NJ, Kumari M, Marmot M, Brunner E, Chambers J, Elliott P, Kooner J, Laan M, Org E, Veldre G, Viigimaa M, Cappuccio FP, Ji C, Iacone, R, Strazzullo P, Moley KH, Cheeseman C. 2008. SLC2A9 is a high-capacity urate transporter in humans. PLoS Medicine 5: 197.12. Caulfield MJ, Munroe PB, OD, Witkowska K, Charchar FJ, Doblado M, Evans S, Eyheramendy S, Onipinlae, Howard P, Shaw-Hawkins S, Dobson RJ, Wallace C, Newhouse SJ, Brown M, Connell JM , Dominiczak, Farrall M, Lathrop GM, Samani NJ, Kumari M, Marmot M, Brunner E, Chambers J, Elliott P, Kooner J, Laan M, Org E, Veldre G, Viigima M, Cappuccio FP, Ji C, Iacone , R, Strazzullo P, Moley KH, Cheeseman C. 2008. SLC2A9 is a high-capacity urate transporter in humans. PLoS Medicine 5: 197.
13. Cederbaum AI. 1991. Microsomal generation of reactive oxygen species and their possible role in alcohol hepatotoxicity. Alcohol and Alcoholism Suppl 1: 291296.13. Cederbaum AI. 1991. Microsomal generation of reactive oxygen species and their possible role in alcohol hepatotoxicity. Alcohol and Alcoholism Suppl 1: 291296.
14. Cha BC, Lee EH. 2007. Antioxidant activities of flavonoids from the leaves of Smilax china Linne. Kor J Pharmacogn 38(1): 31-36. 14. Cha BC, Lee EH. 2007. Antioxidant activities of flavonoids from the leaves of Smilax china linne. Kor J Pharmacogn 38 (1): 31-36.
15. Chena L, Yina H, Lanb Z, Maa S, Zhanga C, Yanga Z, Li P, Linc B. 2011. Anti- hyperuricemic and nephroprotective effects of Smilax china L. J Ethnopharmacology 135: 399405.15. Chena L, Yina H, Lanb Z, Maa S, Zhang C, Yanga Z, Li P, Linc B. 2011. Anti- hyperuricemic and nephroprotective effects of Smilax china L. J Ethnopharmacology 135: 399405.
16. Chen CH, Chan HC, Chang YN, Liu BL, Chen YS. 2000. Effects of bacterial strains on sensory quality of Pu - erh tea in an improved pile-fermentation process. J Sens Stud 24: 534553.16. Chen CH, Chan HC, Chang YN, Liu BL, Chen YS. 2000. Effects of bacterial strains on sensory quality of Pu - erh tea in an improved pile-fermentation process. J Sens Stud 24: 534553.
17. Cheng DS, Hua XL. 2006. Todayresearch of Smilax china. J Chin Med Mater 29: 9093.17. Cheng DS, Hua XL. 2006. Todayresearch of Smilax china . J Chin Med Mater 29: 9093.
18. Chen Y, Wang Q, Li B, Li L, Pan LN, Huang Y. 2008. Study on identification and quality for Smilax china L. of compound gout granules. Asia - Pacific Traditional Medicine 4: 23739.18. Chen Y, Wang Q, Li B, Li L, Pan LN, Huang Y. 2008. Study on identification and quality for Smilax china L. of compound gout granules. Asia - Pacific Traditional Medicine 4: 23739.
19. Choi CH, Song ES, Kim SJ, Kang MH. 2003. Antioxidative activities of Castanea crenata Flos. methanol extracts. Korean J Food Sci Technol 35: 1216-1220.19. Choi CH, Song ES, Kim SJ, Kang MH. 2003. Antioxidative activities of Castanea crenata flos . methanol extracts. Korean J Food Sci Technol 35: 1216-1220.
20. Choi HY. 2004. Antimicrobial effect of ethanol extract of Smilax china leaf. Korean J Sanitation 19: 22-30. 20. Choi HY. 2004. Antimicrobial effect of ethanol extract of Smilax china leaf. Korean J Sanitation 19: 22-30.
21. Clarke SE, Harrell AW, Chenery RJ. 1995. Role of aldehyde oxidase in the in vitro conversion of famciclovir to penciclovir in human liver. Drug Metab Dispos 23: 251254.21. Clarke SE, Harrell AW, Chenery RJ. 1995. Role of aldehyde oxidase in in vitro conversion of famciclovir to penciclovir in human liver. Drug Metab Dispos 23: 251254.
22. Conklin, D., Prough, R. and Bhatanagar, A. 2007. Aldehyde metabolism in the cardiovascular system. Mol Biosyst 3: 136150.22. Conklin, D., Prough, R. and Bhatanagar, A. 2007. Aldehyde metabolism in the cardiovascular system. Mol Biosyst 3: 136150.
23. Critchley DJ, Rance DJ, Beedham C. 1992. Subcellular localisation of guinea pig hepatic molybdenum hydroxylases. Biochem Biophys Res Commun 185(1): 54-59.23. Critchley DJ, Rance DJ, Beedham C. 1992. Subcellular localization of guinea pig hepatic molybdenum hydroxylases. Biochem Biophys Res Commun 185 (1): 54-59.
24. Dambrova M, UhleS, Welch CJ, Wikberg JES. 1998. Identification of an N-hydroxyguanidine reducing activity of xanthine oxidase. Eur J Biochem 257: 178184.24. Dambrova M, Uhle S, Welch CJ, Wikberg JES. 1998. Identification of an N-hydroxyguanidine reducing activity of xanthine oxidase. Eur J Biochem 257: 178184.
25. Das SK, Nayak P, Vasudevan DM. 2005. Consequences of ethanol consumption. J Indian Soc Toxicol 1: 110.25. Das SK, Nayak P, Vasudevan DM. 2005. Consequences of ethanol consumption. J Indian Soc Toxicol 1: 110.
26. De Haan JB, Cristiano F, Iannello RC, Kola I (1995) Cu/Zn-superoxide dismutase and glutathione peroxidase during aging. Biochem Mol Biol Int 35: 1281-1297. 26. De Haan JB, Cristiano F, Iannello RC, Kola I (1995) Cu / Zn-superoxide dismutase and glutathione peroxidase during aging. Biochem Mol Biol Int 35: 1281-1297.
27. Ellman GL (1959) Tissue sulfhydryl group. Arch Biochem Biophys 82: 7077.27. Ellman GL (1959) Tissue sulfhydryl group. Arch Biochem Biophys 82: 7077.
28. Franke WW, Denk H, Schmid E, Osborn M, and Weber K. 1979. Ultrastructural, biochemical and immunological characterization of Mallory bodies in livers of griseofulvin-treated mice: Fimbriated rods of filaments containing prekeratin- like polypeptides. Lab Invest 40: 207220.28. Franke WW, Denk H, Schmid E., Osborn M, and Weber K. 1979. Ultrastructural, biochemical and immunological characterization of Mallory bodies in livers of griseofulvin-treated mice: Fimbriated rods of filaments containing prekeratin-like polypeptides. Lab Invest 40: 207220.
29. Friedewald WT, Levy RI, and Fredrickson DS (1972) Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge. Clin Chem 18: 499502.29. Friedewald WT, Levy RI, and Fredrickson DS (1972) Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without the use of the preparative ultracentrifuge. Clin Chem 18: 499502.
30. Frings CS, Fendley TW, Dunn RT, Queen CA. 1972. Improved determination of total serum lipids by the sulfo-phospho-vanillin reaction. Clin Chem 18: 673-674.30. Frings CS, Fendley TW, Dunn RT, Queen CA. 1972. Improved determination of total serum lipids by the sulfo-phospho-vanillin reaction. Clin Chem 18: 673-674.
31. Garattini E, Fratelli M, Terao M. 2008. Mammalian aldehyde oxidases: genetics, evolution and biochemistry. Cell Mol Life Sci 65: 10191048.31. Garattini E, Fratelli M, Terao M. 2008. Mammalian aldehyde oxidases: genetics, evolution and biochemistry. Cell Mol Life Sci 65: 10191048.
32. Garattini E, Terao M. 2011. Increasing recognition of the importance of aldehyde oxidase in drug development and discovery. Drug Metab Rev 43: 374386.32. Garattini E, Terao M. 2011. Increasing recognition of the importance of aldehyde oxidase in drug development and discovery. Drug Metab Rev 43: 374386.
33. Garcia-Conesa MT, P, Kauppinen S, Williamson G. 2001. Hydrolysis of diethyl diferulates by a tannase from Aspergillus oryzae . Carbohydrate Polymers 44: 319324.33. Garcia-Conesa MT, P, Kauppinen S, Williamson G. 2001. Hydrolysis of diethyl diferulates by a tannase from Aspergillus oryzae. Carbohydrate Polymers 44: 319324.
34. Gruchow HW, Sobocinski KA Barboriac JJ, Scheller JG. 1985. Alcohol consumption, nutrient intake and relative body weight among US adults . Am Clin Nutr 42: 289-595.34. Gruchow HW, Sobocinski KA Barboriac JJ, Scheller JG. 1985. Alcohol consumption, nutrient intake and relative body weight among US adults . Am Clin Nutr 42: 289-595.
35. Halliwell B. 2006. Reactive oxygen species and the central nervous system. J Neurochem 59: 1609-1623. 35. Halliwell B. 2006. Reactive oxygen species and the central nervous system. J Neurochem 59: 1609-1623.
36. Ham YK and Kim SW. 2004. Protective effects of plant extract on the hepatocytes of rat treated with carbon tetrachloride. J Korean Soc Food Sci Nutr 33: 12461251.36. Ham YK and Kim SW. 2004. Protective effects of plant extracts on hepatocytes of rat treated with carbon tetrachloride. J Korean Soc Food Sci Nutr 33: 12461251.
37. Hamzeh-Mivehroud M, Rahmani S, Rashidi MR, Hosseinpour Feizi MA, Dastmalchi S. 2013. Structure-based investigation of rat aldehyde oxidase inhibition by flavonoids. Xenobiotica Jan 3.37. Hamzeh-Mivehroud M, Rahmani S, Rashidi MR, Hosseinpour Feizi MA, Dastmalchi S. 2013. Structure-based investigation of rat aldehyde oxidase inhibition by flavonoids.
38. Herbert A, Jeol LS. 1993. Sensory evaluation practices. 2nd ed. Academic Press, New York, USA. p 68-94. 38. Herbert, Jeol LS. 1993. Sensory evaluation practices. 2nd ed. Academic Press, New York, USA. p 68-94.
39. Hirao Y, Kitamura S, Tatsumi, K. 1994. Epoxide reductase activity of mammalian liver cytosols and aldehyde oxidase. Carcinogenesis 15: 739743.39. Hirao Y, Kitamura S, Tatsumi, K. 1994. Epoxide reductase activity of mammalian liver cytosols and aldehyde oxidase. Carcinogenesis 15: 739743.
40. Holasova, M., Fiedlerova, V., Smrcinova, H., Orsak, M., Lachman, J. and Vavreinova, S. 2002. Buckwheat the source of antioxidant activity in functional foods. Food Res Int 35: 207-211.40. Holasova, M., Fiedlerova, V., Smrcinova, H., Orsak, M., Lachman, J. and Vavreinova, S. 2002. Buckwheat the source of antioxidant activity in functional foods. Food Res Int 35: 207-211.
41. Horiuchi H, Ota M, Nishimura S, Kaneko H, Kasahara Y, Ohta T, Komoriya K. 2000. Allopurinol induces renal toxicity by impairing pyrimidine metabolism in mice. Life Sci14; 66(21): 2051-2070.41. Horiuchi H, Ota M, Nishimura S, Kaneko H, Kasahara Y, Ohta T, Komoriya K. 2000. Allopurinol induces renal toxicity by impairing pyrimidine metabolism in mice. Life Sci 14; 66 (21): 2051-2070.
42. Huang DY. and Ichikawa Y. 1994. Two different enzymes are primarily responsible for retinoic acid synthesis in rabbit liver cytosol. Biochem Biophys Res Commun 205: 12781283. 42. Huang DY. and Ichikawa Y. 1994. Two different enzymes are primarily responsible for retinoic acid synthesis in rabbit liver cytosol. Biochem Biophys Res Commun 205: 12781283.
43. Hur SJ, Ye BW, Lee JL, Ha YL, Park GB, Joo ST. 2004. Effect of conjugated linoleic acid on color and lipid oxidation of beef patties during cold storage. Meat Sci 66: 771-775.43. Hur SJ, Ye BW, Lee JL, Ha YL, Park GB, Joo ST. 2004. Effect of conjugated linoleic acid on color and lipid oxidation of beef patties during cold storage. Meat Sci 66: 771-775.
44. Itoh K. 2009. Individual and strain differences of aldehyde oxidase in the rat. Yakugaku Zasshi 129: 1487-1493. 44. Itoh K. 2009. Individual and strain differences of aldehyde oxidase in the rat. Yakugaku Zasshi 129: 1487-1493 .
45. Johanna W, Markus N, Sabrina B, Wolfgang M, Andreas A, Aiman O, Benedikt G, Arndt H, Andreas S, Charalarmpos A, Jurgen S, Christa B. 2008. Small-interference RNA-mediated knock-down of aldehyde oxidase 1 in 3T3-L1 cells impairs adipogenesis and adiponectin release. FEBS letters 582: 2965-2972.45. Johanna W, Markus N, Sabrina B, Wolfgang M, Andreas A, Aiman O, Benedikt G, Arndt H, Andreas S, Charalarmpose, Jurgen S, Christa B. 2008. Small-interference RNA-mediated knockdown of
46. Jordan CGM, Rashidi MR, Laljee H, Clarke SE, Brown JE, Beedham C. 1999. Aldehyde oxidase-catalyzed oxidation of methotrexate in the liver of guinea- pig, rabbit and man. J Pharm Pharmacol 51: 411-418. 46. Jordan CGM, Rashidi MR, Laljee H, Clarke SE, Brown JE, Beedham C. 1999. Aldehyde oxidase-catalyzed oxidation of methotrexate in the liver of guinea pigs, rabbit and man. J Pharm Pharmacol 51: 411-418.
47. Kang YH, Park YK, Oh SR, Moon KD. 1995. Studies on the physiological functionality of pine needle and mugwort extracts. Korean J Food Sci Technol 27: 978-984. 47. Kang YH, Park YK, Oh SR, Moon KD. 1995. Studies on the physiological function of pine needle and mugwort extracts. Korean J Food Sci Technol 27: 978-984.
48. Kawashima K, Hosoi K, Naruke T, Shiba T, Kitamura M, Watabe T. 1999. Aldehyde oxidasedependent marked species difference in hepatic metabolism of the sedativehypnotic, zeleplon, between monkeys and rats. Drug Metab Dispos 27: 422-428. 48. Kawashima K, Hosoi K, Naruke T, Shiba T, Kitamura M, Watabe T. 1999. Aldehyde oxidisedependent marked species differences in the sedative hypnotics, zeleplon, between monkeys and rats. Drug Metab Dispos 27: 422-428.
49. Kitamura S, Nakatani K, Sugihara K, Ohta S. 1999. Strain differences of the ability to hydroxylate methotrexate in rats. Comp Biochem Physiol 122C: 331-336. 49. Kitamura S, Nakatani K, Sugihara K, Ohta S. 1999. Strain differences in the ability to hydroxylate methotrexate in rats. Comp Biochem Physiol 122C: 331-336.
50. Ko MS, Yang JB. 2011. Antioxidant and antimicrobial activities of Smilax china leaf extracts. Korean J Food Preserv 18(5): 764-772.50. Ko MS, Yang JB. 2011. Antioxidant and antimicrobial activities of Smilax chin a leaf extracts. Korean J Food Preserv 18 (5): 764-772.
51. Krenitsky TA (1978) Aldehyde oxidase and xanthine oxidase-functional and evolutionary relationships. Biochem Pharmacol 27: 2763-2764.51. Krenitsky TA (1978) Aldehyde oxidase and xanthine oxidase-functional and evolutionary relationships. Biochem Pharmacol 27: 2763-2764.
52. Kundu TK, Hille R, Velayutham M, Zweier JL. 2007. Characterization of superoxide production from aldehyde oxidase: an important source of oxidants in biological tissues. Arch Biochem Biophys 460: 113-121.52. Kundu TK, Hille R, Velayutham M, Zweier JL. 2007. Characterization of superoxide production from aldehyde oxidase: an important source of oxidants in biological tissues. Arch Biochem Biophys 460: 113-121.
53. Lee YK, Lee SI, Kim JS, Yang SH, Lee IA, Kim SD, Suh JW. 2012. Antioxidant activity of green tea fermented with Monascus pilosus. J Appl Biol Chem 55(1): 19-25.53. Lee YK, Lee SI, Kim JS, Yang SH, Lee IA, Kim SD, Suh JW. 2012. Antioxidant activity of green tea fermented with Monascus pilosus J Appl Biol Chem 55 (1): 19-25.
54. Lieber CS. 1991. Hepatic, metabolic and toxic effects of ethanol. Alcohol Clin Exp Res 15: 573-592.54. Lieber CS. 1991. Hepatic, metabolic and toxic effects of ethanol. Alcohol Clin Exp Res 15: 573-592.
55. Lieber CS. 1992. Medical and Nutritional Complications of Alcoholism: Mechanisms and Management. New York, Plenum Press.55. Lieber CS. 1992. Medical and Nutritional Complications of Alcoholism: Mechanisms and Management. New York, Plenum Press.
56. Lieber CS. 1997. Ethanol metabolism, cirrhosis and alcoholism. Clinica Chimica Acta 257: 59-84.56. Lieber CS. 1997. Ethanol metabolism, cirrhosis and alcoholism. Clinica Chimica Acta 257: 59-84.
57. Lieber CS. 2008. Alcoholic fatty liver : its pathogenesis and mechanism of progression to inflammation and fibrosis. Alcohol 34:9-19 2004; Lu Y, Ce derbaum AI: CYP2E1 and oxidative liver injury by alcohol. Free Radic Biol Med 44: 723-738.57. Lieber CS. 2008. Alcoholic fatty liver: its pathogenesis and mechanism of progression to inflammation and fibrosis. Alcohol 34: 9-19 2004; Lu Y, Ce derbaum AI: CYP2E1 and oxidative liver injury by alcohol. Free Radic Biol Med 44: 723-738.
58. Li YL, Gan GP, Zhang HZ, Wu HZ, Li CL, Huang YP, Liu YW, Liu JW. 2007. A flavonoid glycoside isolated from Smilax china L. rhizome in vitro anticancer effects on human cancer cell lines. J Ethnopharmacol 113: 115124.58. Li YL, Gan GP, Zhang HZ, Wu HZ, Li CL, Huang YP, Liu YW, Liu JW. 2007. A flavonoid glycoside isolated from Smilax china L. rhizome in vitro anticancer effects on human cancer cell lines. J Ethnopharmacol 113: 115124.
59. Lowry OH, Rosebrough NJ, Farr AL, and Randall RL (1951) Protein measurement by folin phenol reagent. J Biol Chem 193: 265275.59. Lowry OH, Rosebrough NJ, Farr AL, and Randall RL (1951) Protein measurement by folin phenol reagent. J Biol Chem 193: 265275.
60. Luciana GDO, Francisca MB, Cybelle RDL. 2009. Effects of different concentrations of sugarcane alcohol on food intake and nutritional status of male and female periadolescent rats Alcohol 43: 137-146.60. Luciana GMO, Francisca MB, Cybelle RDL. 2009. Effects of different concentrations of sugarcane alcohol on food intake and nutritional status of male and female periadolescent rats Alcohol 43: 137-146.
61. Maia L, Mira L. 2002. Xanthine oxidase and aldehyde oxidase: A simple procedure for the simultaneous purification from rat liver. Arch Biochem Biophys 400: 48-53. 61. Maia L, Mira L. 2002. Xanthine oxidase and aldehyde oxidase: A simple procedure for simultaneous purification from rat liver. Arch Biochem Biophys 400: 48-53.
62. McCrystal M, Evans B, Harvey V, Thompson P, Porter D, Baguley B. 1999. Phase 1 study of the cytotoxic agent N-w2-(dimethylamino)ethylxacridine-4- carboxamide. Cancer Chemother Pharmacol 44: 3944.62. McCrystal M, Evans B, Harvey V, Thompson P, Porter D, Baguley B. 1999.
63. Meda A, Lamien CE, Romito M, Millogo J, Nacoulma OG. 2005. Determination of the total phenolic, flavonoid and proline contents in burkina fasan honey, as well as their radical scavenging activity. Food Chem 91: 571577. 63. Medae, Lamien CE, Romito M, Millogo J, Nacoulma OG. 2005. Determination of the total phenolic, flavonoid and proline contents of burkina fasan honey, as well as their radical scavenging activity. Food Chem 91: 571577.
64. Mercader J, Ribot J, Murano I, Felipe F, Cinti S, Bonet ML. and Palou A. 2006. Remodeling of white adipose tissue after retinoic acid administration in mice. Endocrinology 147: 53255332.64. Mercader J, Ribot J, Murano I, Felipe F, Cinti S, Bonet ML. and Palou A. 2006. Remodeling of white adipose tissue after retinoic acid administration in mice. Endocrinology 147: 53255332.
65. Minussi RC, Rossi M, Bologna L, Cordi L, Rotilio D, Pastore GM, Duran N. 2003. Phenolic compounds and total antioxidant potential of commercial wines. Food Chem 82: 409416. 65. Minussi RC, Rossi M, Bologna L, Cordi L, Rotilio D, Pastore GM, Duran N. 2003. Phenolic compounds and total antioxidant potential of commercial wines. Food Chem 82: 409416.
66. Mira L, Maia L, Barreira L, Manso CF. 1995. Evidence for free radical generation due to NADH oxidation by aldehyde oxidase during ethanol metabolism. Arch Biochem Biophys 318: 53-58.66. Mira L, Maia L, Barreira L, Manso CF. 1995. Evidence for free radical generation due to NADH oxidation by aldehyde oxidase during ethanol metabolism. Arch Biochem Biophys 318: 53-58.
67. Moon SH, Lee MK, Chae KS. 2001. Inhibitory effects of the solvent fractions from persimmon leaves on xanthine oxidase activity. Kor J Food Nutr 14: 120-125.67. Moon SH, Lee MK, Chae KS. 2001. Inhibitory effects of the solvent fractions from persimmon leaves on xanthine oxidase activity. Kor J Food Nutr 14: 120-125.
68. Moriwaki Y, Yamamoto T, Nasako Y, Takahashi S, Suda M, Hiroishi K, Hada T, Higashino K. 1993. In vitro oxidation of pyrazinamide and allopurinol by rat liver aldehyde oxidase. Biochem Pharmacol 46: 975-981. 68. Moriwaki Y, Yamamoto T, Nasako Y, Takahashi S, Suda M, Hiroishi K, Hada T, Higashino K. 1993. In vitro oxidation of pyrazinamide and allopurinol by rat liver aldehyde oxidase. Biochem Pharmacol 46: 975-981.
69. Neumeier M, Weigert J, SchA, Weiss TS, Schmidl C, BR, Bollheimer C, Aslanidis C, SchJ, Buechler C. 2006. Aldehyde oxidase 1 is highly abundant in hepatic steatosis and is downregulated by adiponectin and fenofibric acid in hepatocytes in vitro. Biochem Biophys Res Commun 350: 731-735.69. Neumeier M, Weigert J, Scha, Weiss TS, Schmidl C, BR, Bollheimer C, Aslanidis C, SchJ, Buechler C. 2006.
70. Nishino T, Tamura I. 1991. The mechanisim of conversion of xanthine dehydrogenase to oxidase and therole of the enzyme in the resperfusion injury. Adv Exp Med Biol 309: 327-337. 70. Nishino T, Tamura I. 1991. The mechanism of conversion of xanthine dehydrogenase to oxidase and the enzyme of the reserfusion injury. Adv Exp Med Biol 309: 327-337.
71. Oei HH, Kentroo WE, Burton KP, and Schaffer SW. 1982. A possible role of xanthine oxidase in pproducing oxidative stress in the heart of chronically ethanol treated rats. Res Commun Chem Pathol Pharmacol 38: 453461.71. Oei HH, Kentroo WE, Burton KP, and Schaffer SW. 1982. A possible role of xanthine oxidase in the production of oxidative stress in the heart of chronically ethanol treated rats. Res Commun Chem Pathol Pharmacol 38: 453461.
72. Osawa T. 1994. Novel natural antioxidant for utilization in food and biological system. In Postharvest Biochemistry of Plant Food Material in the Tropics. Uritani I, Garcia VV, Mendoza EM, eds. Japan Scientific Societies Press, Tokyo, Japan. p. 241-251.72. Osawa T. 1994. Novel natural antioxidants for utilization in food and biological systems. In Postharvest Biochemistry of Plant Food Material in the Tropics . Uritani I, Garcia VV, Mendoza EM, eds. Japan Scientific Societies Press, Tokyo, Japan. p. 241-251.
73. Owuor PO, Obanda M, Nyirenda HE, Mphangwe NIK, Wright LP, Apostolides Z. 2006. The relationship between some chemical parameters and sensory evaluations for plain black tea (Camellia sinensis) produced in Kenya and comparison with similar teas from Malawi and South Africa. Food Chem 97: 644-653. 73. Owuor PO, Obanda M, Nyirenda HE, Mphangwe NIK, Wright LP, Apostolides Z. 2006. The relationship between some chemical parameters and sensory evaluations for plain black tea ( Camellia sinensis ) produced in Kenya and comparison with similar teas from Malawi and South Africa. Food Chem 97: 644-653.
74. Pikaar NA, Wedel M, Vander BEJ, Van DW and Hermus RJ. 1987. Effects of moderate alcohol consumption on platelet aggregation fibrinolysis and blood lipids. Metabolism 36: 538-548.74. Pikaar NA, Wedel M, Vander BEJ, Van DW and Hermus RJ. 1987. Effects of moderate alcohol consumption on platelet aggregation fibrinolysis and blood lipids. Metabolism 36: 538-548.
Pryde DC, Dalvie D, Hu Q, Jones P, Obach RS, Tran TD. 2010. Aldehyde oxidase: an enzyme of emerging importance in drug discovery. J Med Chem 53: 84418460.Pryde DC, Dalvie D, Hu Q, Jones P, Obach RS, Tran TD. 2010. Aldehyde oxidase: an enzyme of emerging importance in drug discovery. J Med Chem 53: 84418460.
75. Rajagopalan KV, Fridovich I, Handler P. 1962. Hepatic aldehyde oxidase. . Purification and properties. J Biol Chem 237: 922-928.75. Rajagopalan KV, Fridovich I, Handler P. 1962. Hepatic aldehyde oxidase. . Purification and properties. J Biol Chem 237: 922-928.
76. Ruan HL, Zhang YH, Zhao W, Tan YF, Sun ZL, Wu JZ. 2002. Studies on the chemical constituents of Smilax china L. Natural Product Research and Development 14: 3536.76. Ruan HL, Zhang YH, Zhao W, Tan YF, Sun ZL, Wu JZ. 2002. Studies on the chemical constituents of Smilax china L. Natural Product Research and Development 14: 3536.
77. Rashidi MR, Nazemiyeh H. 2010. Inhibitory effects of flavonoids on molybdenum hydroxylases activity. Expert Opin Drug Metab Toxicol 6(2): 133-152.77. Rashidi MR, Nazemiyeh H. 2010. Inhibitory effects of flavonoids on molybdenum hydroxylases activity. Expert Opin Drug Metab Toxicol 6 (2): 133-152.
78. Robert A, Terkeltaub, M.D. 2003. Gout. N Engl J Med ;349:1647-1655.78. Roberta, Terkeltaub, MD 2003. Gout. N Engl J Med ; 349: 1647-1655.
Salmela KS, Kessova IG, Tsyrolv IB, Lieber CS. 1998. Respective roles of cytochrome P450 2E1, 1A2, and 3A4 in the hepatic microsomal ethanol oxidizing system. Alcoholism , Clinical and Experimental Research 22: 21252132.Salmela KS, Kessova IG, Tsyrolv IB, Lieber CS. 1998. Respective roles of cytochrome P450 2E1, 1A2, and 3A4 in the hepatic microsomal ethanol oxidizing system. Alcoholism , Clinical and Experimental Research 22: 21252132.
79. Shaw S, Jayatilleke E. 1990. The physiological role of aldehyde oxidase in ethanol- induced hepatic lipid peroxidation in the rat. Biochem J. 268(3): 579-583.79. Shaw S, Jayatilleke E. 1990. The physiological role of aldehyde oxidase in ethanol-induced hepatic lipid peroxidation in the rat. Biochem J. 268 (3): 579-583.
80. Shaw S, Jayatilleke E. 1990. The role of aldehyde oxidase in ethanol-induced hepatic lipid peroxidation in the rat. Biochem J 268: 579583.80. Shaw S, Jayatilleke E. 1990. The role of aldehyde oxidase in ethanol-induced hepatic lipid peroxidation in the rat. Biochem J 268: 579583.
81. Shu XS, Gao ZH, Yang XL. 2006. Anti-inflammatory and anti-nociceptive activities of Smilax china L. aqueous extract. J Ethnopharmacol 103: 32732. 81. Shu XS, Gao ZH, Yang XL. 2006. Anti-inflammatory and anti-nociceptive activities of Smilax china L. aqueous extract. J Ethnopharmacol 103: 32732.
82. Song HS, Park YH, Jung SH, Kim DP, Jung YH, Lee MK, Moon KY. 2006. Antioxidant activity of extracts from Smilax china root. J Korean Soc Food Sci Nutr 35(9): 1133-1138. 82. Song HS, Park YH, Jung SH, Kim DP, Jung YH, Lee MK, Moon KY. 2006. Antioxidant activity of extracts from Smilax china root. J Korean Soc Food Sci Nutr 35 (9): 1133-1138.
83. Stirpe F, Della Corte E. 1969. The regulation of rat liver xanthine oxidase. Conversion in vitro of the enzyme activity from dehydrogenase (type D) to oxidase (type O). J Biol Chem 244: 3855-3863.83. Stirpe F, Della Corte E. 1969. The regulation of rat liver xanthine oxidase. Conversion in vitro of enzyme activity from dehydrogenase (type D) to oxidase (type O). J Biol Chem 244: 3855-3863.
84. Stoddart AM, Levine WG. 1992. Azoreductase activity by purified rabbit liver aldehyde oxidase Biochem Pharmacol 43: 2227-2235.84. Stoddart AM, Levine WG. 1992. Azoreductase activity by purified rabbit liver aldehyde oxidase Biochem Pharmacol 43: 2227-2235.
85. Sugihara K, Kitamura S, Tatsumi K. 1996. Involvement of mammalian liver cytosols and aldehyde oxidase in reductive metabolism of zonisamide. Drug Metab Dispos 24: 199-202. 85. Sugihara K, Kitamura S, Tatsumi K. 1996. Involvement of mammalian liver cytosols and aldehyde oxidase in reductive metabolism of zonisamide. Drug Metab Dispos 24: 199-202.
86. Lee SW, Kim KS, Kim SD. 1974. Studies on the compositional changes of pomegranate fruit during maturity. II. Changes in polyphenol compounds and anthocyan pigments. J Korean Soc Hort Sci 15(1) 64-67. 86. Lee SW, Kim KS, Kim SD. 1974. Studies on the compositional changes of pomegranate fruit during maturity. II. Changes in polyphenol compounds and anthocyan pigments. J Korean Soc Hort Sci 15 (1) 64-67.
87. Tatsumi K, Kitamura S, Yamada H. 1983. Sulfoxide reductase activity of liver aldehyde oxidase. Biochim Biophys Acta 747: 86-92.87. Tatsumi K, Kitamura S, Yamada H. 1983. Sulfoxide reductase activity of liver aldehyde oxidase. Biochim Biophys Acta 747: 86-92.
88. Tatsumi K, Kitamura S, Narai N. 1986. Reductive metabolism of aromatic nitro compounds including carcinogens by rabbit liver preparations. Cancer Res 46: 10891093.88. Tatsumi K, Kitamura S, Narai N. 1986. Reductive metabolism of aromatic nitro compounds including carcinogens by rabbit liver preparations. Cancer Res 46: 10891093.
89. Torel J, Gillard J, Gillard P. 1986. Antioxidant activity of flavonoids and reactivity with peroxy radical. Phytochem 25: 383-385.89. Torel J, Gillard J, Gillard P. 1986. Antioxidant activity of flavonoids and reactivity with peroxy radicals. Phytochem 25: 383-385.
90. Urano S, Midori H, Tochihi N, Matsuo M, Shiraki M, and Ito H. 1991. Vitamin E and the susceptibility of erythrocytes and reconstituted liposome to oxidative stress in aged diabetics. Lipids 26: 5862.90. Urano S, Midori H, Tochihi N, Matsuo M, Shiraki M, and Ito H. 1991. Vitamin E and the susceptibility of erythrocytes and reconstituted liposomes to oxidative stress in aged diabetics. Lipids 26: 5862.
91. Vohl MC, Sladek R, Robitaille J, Gurd S, Marceau P, Richard D, Hudson TJ. and Tchernof A. 2004. A survey of genes differentially expressed in subcutaneous and visceral adipose tissue in men. Obes Res 12: 12171222.91. Vohl MC, Sladek R, Robitaille J, Gurd S, Marceau P, Richard D, Hudson TJ. and Tchernof A. 2004. A survey of genes differentially expressed in subcutaneous and visceral adipose tissue in men. Obes Res 12: 12171222.
92. Weigert J, Neumeier M, Bauer S, Mages W, Schnitzbauer AA, Obed A, GrB, Hartmann A, SchA, Aslanidis C, SchJ, Buechler C. 2008. Small-interference RNA-mediated knock-down of aldehyde oxidase 1 in 3T3-L1 cells impairs adipogenesis and adiponectin release. FEBS Lett 582: 2965-2972. Small-interfering RNA-mediated knockdown of aldehyde oxidase 1 (IGF-1) is a key component of this process. in 3T3-L1 cells impairs adipogenesis and adiponectin release. FEBS Lett 582: 2965-2972.
93. Wortmann RL. 2002. Gout and hyperuricemia. Current Opinion in Rheumatology 14: 281286.93. Wortmann RL. 2002. Gout and hyperuricemia. Current Opinion in Rheumatology 14: 281286.
94. Wu LS, Wang, XJ, Wang H, Yang HW, Jia AQ, Ding Q. 2010. Cytotoxic polyphenols against breast tumor cell in Smilax china L. J Ethnopharmacol 130: 46064.94. Wu LS, Wang, XJ, Wang H, Yang HW, Jia AQ, Ding Q. 2010. Cytotoxic polyphenols against breast tumor cell in Smilax china L. J Ethnopharmacol 130: 46064.
95. Xu Y, Liang JY, Zou ZM. 2008. Studies on chemical constituents of rhizomes of Smilax china L. China J Chinese Materia Medica 21: 119121.95. Xu Y, Liang JY, Zou ZM. 2008. Studies on chemical constituents of rhizomes of Smilax China L. China J Chinese Materia Medica 21: 119121.
96. Yamaguchi Y, Matsumura T, Ichida K, Okamoto K, Nishino T. 2007. Human xanthine oxidase changes its substrate specificity to aldehyde oxidase type upon mutation of amino acid residues in the active site: roles of active site residues in binding and activation of purine substrate. J Biochem 141: 513-524.96. Yamaguchi Y, Matsumura T, Ichida K, Okamoto K, Nishino T. 2007. Human xanthine oxidase changes its substrate specificity to aldehyde oxidase type upon mutation of amino acid residues in the active site: of purine substrate. J Biochem 141: 513-524.
97. Yasuhara A, Akiba-Goto M, Fujishiro K, Uchida H, Uwajima T, Aisaka K. 2002. Production of aldehyde oxidases by microorganisms and their enzymatic properties. J Biosci Bioengineering 94(2): 124-129. 97. Yasuharaa, Akiba-Goto M, Fujishiro K, Uchida H, Uwajima T, Aisaka K. 2002. Production of aldehyde oxidases by microorganisms and their enzymatic properties. J Biosci Bioengineering 94 (2): 124-129.
98. Zhu JX, Wang Y, Kong LD, Yang C, Zhang X. 2004. Effects of Biota orientalis extract and its flavonoid constituents, quercetin and rutin on serum uric acid levels in oxonate-induced mice and xanthine dehydrogenase and xanthine oxidase activities in mouse liver. J Ethnopharm 93: 133-140.98. Zhu JX, Wang Y, Kong LD, Yang C, Zhang X. 2004. Effects of Biota orientalis extract and its flavonoid constituents, quercetin and rutin on serum uric acid levels in oxonate-induced mice and xanthine dehydrogenase and xanthine oxidase activities in mouse liver. J Ethnopharm 93: 133-140.
Claims (20)
상기 발효에 의해 발효 전보다 향상된 잔틴 옥시다아제(Xanthine oxidase) 억제 활성 또는 알데하이드 옥시다아제(aldehyde oxidase) 억제 활성을 나타내는 것을 특징으로 하는 발효 청미래덩굴 잎.
It is a product obtained by fermenting the leaf of Vinegar with the Aspergillus microorganism,
Wherein the fermented green leaf is characterized by exhibiting an activity of inhibiting xanthine oxidase or an activity of inhibiting aldehyde oxidase when fermented.
The method of claim 1, wherein the microorganism is Aspergillus seusok Aspergillus Kawachi (Aspergillus kawachii , Aspergillus awamori , Aspergillus oryzae ), aspergillus ( Aspergillus sojae) or Aspergillus fermentation Smilax China leaf, characterized in that consisting of at least one selected from Usami (Aspergillus usamii).
The aspergillus oryzae according to claim 2, wherein the Aspergillus oryzae is Aspergillus oryzae KACC 40247 strain, Aspergillus oryzae KFRI 995 strain, Aspergillus oryzae var. brunneus KACC 44823 strain or Aspergillus oryzae var effuses characterized by consisting of one or more selected from the KACC 44990 strains.
The fermented blue-violet creeper blade according to claim 1, wherein the fermented Cynthia sinensis leaf is in powder form.
[Claim 5] The fermented blueberry leaf of claim 4, wherein the powder has a particle size of 20 to 100 mesh.
(b) 발효된 청미래덩굴 잎을 건조하는 단계를 포함하고,
상기 발효된 청미래덩굴 잎은 발효 전보다 향상된 잔틴 옥시다아제(Xanthine oxidase) 억제 활성 또는 알데하이드 옥시다아제(aldehyde oxidase) 억제 활성을 나타내는 것을 특징으로 하는 발효 청미래덩굴 잎의 제조방법.
(a) inoculating an Aspergillus microflora culture solution to the leaves of Cheongmyeongjae and fermenting the Cheongmyeonghwa leaves; And
(b) drying the fermented blue-green vine leaves,
Wherein the fermented Cyanoma vine leaves exhibit an activity of inhibiting xanthine oxidase or an activity of inhibiting aldehyde oxidase when compared with that before fermentation.
The microorganism according to claim 6, wherein the aspergillus microorganism is Aspergillus kawachii , Aspergillus awamori , Aspergillus oryzae ), aspergillus ( Aspergillus sojae) or Aspergillus Usami (Method of producing fermentation Smilax China leaf, characterized in that consisting of at least one selected from Aspergillus usamii).
The aspergillus oryzae according to claim 7, wherein the Aspergillus oryzae is Aspergillus oryzae KACC 40247 strain, Aspergillus oryzae KFRI 995 strain, Aspergillus oryzae var. brunneus KACC 44823 strain or Aspergillus oryzae var effuses KACC 44990 strain. < RTI ID = 0.0 > 18. < / RTI >
[7] The method according to claim 6, further comprising a step of drying and crushing the Cheongmaia crane leaf before the step (a).
[10] The method according to claim 9, wherein the Rhododendron vine leaf is pulverized into powder having a particle size of 20 to 100 mesh.
[10] The method according to claim 10, further comprising the step of sterilizing the powder of Cheongmyeonghwa blade before the step (a).
[Claim 7] The method according to claim 6, wherein the microorganism concentration of the aspherulis microorganism culture is 10 6 cells / ml to 10 10 cells / ml, and the inoculum amount is 1 to 15% ≪ / RTI >
[Claim 7] The method according to claim 6, wherein the Rhododendron vine leaf is fermented in a solid state at a total water content of 50 to 75%.
7. The method according to claim 6, wherein the fermentation temperature is 20 to 35 DEG C and the fermentation time is 5 to 30 days.
[7] The method according to claim 6, further comprising the step of sterilizing the fermented Vine leaves after fermentation between step (a) and step (b).
A fermented tea comprising a fermented green tea leaf of any one of claims 1 to 5 or a fermented green tea leaf produced by the method of any one of claims 6 to 15.
17. The fermented tea according to claim 16, wherein the fermented tea is composed of a tea bag and a fermented blue-green vine leaf contained therein.
[17] The fermented tea according to claim 16, wherein the fermentation tea further comprises at least one selected from corn root powder, corn stalk powder, gherkin powder or ganoderma powder.
A functional beverage comprising a fermented blueberry blade according to any one of claims 1 to 5 or a water extract of fermented bluefin tangerine leaves produced by the method according to any one of claims 6 to 15.
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2007284397A (en) | 2006-04-18 | 2007-11-01 | Toshie Yasuda | Method for producing functional composition |
KR20110076043A (en) * | 2009-12-29 | 2011-07-06 | 주식회사 코리아나화장품 | Cosmetic composition comprising smilax china as active ingredient |
KR20120029115A (en) * | 2010-09-16 | 2012-03-26 | 김도연 | Composition for heavy metal detoxification |
KR20120029117A (en) * | 2010-09-16 | 2012-03-26 | 김도연 | Composition for promoting nicotine degradation |
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KR101246266B1 (en) * | 2010-11-24 | 2013-03-21 | 동아대학교 산학협력단 | Method for preparing fermented silkworm powder and composition comprising fermented silkworm powder for the preventing or treating of hyperlipidemia and fatty liver |
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KR20110076043A (en) * | 2009-12-29 | 2011-07-06 | 주식회사 코리아나화장품 | Cosmetic composition comprising smilax china as active ingredient |
KR20120029115A (en) * | 2010-09-16 | 2012-03-26 | 김도연 | Composition for heavy metal detoxification |
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108850366A (en) * | 2018-08-08 | 2018-11-23 | 广州宏韵医药科技股份有限公司 | A kind of alternative tea composition and preparation method thereof with anti-trioxypurine effect |
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