본 연구에서는 해삼속의 폴리페놀과 플라보노이드의 생리활성물질을 추출하기 위한 용매분획의 수율을 확인하고자 연구를 시도하였다. 이미 보고된 사례에서 50%에탄올 추출 용매분획은 해삼 항산화 물질의 높은 수율 결과로 확인되었다. 해삼의 항산화물질 추출량을 결정짓는 것은 추출에 적용된 추출용 매분획의 결과로 확인되었다. 또한 ‘추출용매에 따라 해삼생리활성 항산화물질 추출함량이 크게 차이가 있다.’는 선행연구를 통해 용매분획추출에 관한 필요성의 결과를 얻었다. 50%에탄올 추출 용매분획의 해삼추 출물에 포함된 페놀물질의 높은 항산화성분 추출 결과가 증명되었다. 추출용매분획 연구사례에서 수율이 가장 저조한 아세트산에틸 용매분획은 다른 용매분획에 비해 높은 페놀함량을 수확하여 항산화효과가 확인되었다. 이에, 추출 용매분획물의 적용에 따른 수율변화를 통해 항산화추출물의 높은 수율에 미치는 영향을 확인 하였다. 따라서 본 연구를 통해 50%에탄올 용매의 최적화된 해삼생리활성물질 추출 용매분획으로 검증되었다.

Styela clava (SC), a sea squirt has a biphasic life cycle, showed the animal phase during larval life and the vegetable phase during adult life. The adult SC becomes fixed on hard surface and covered with thick cellulose membrane to protect itself from various physical and chemical damages in marine environment. Especially, adult SC forms a huge cystic space as a portion of digestive tract, filled with seawater containing various planktons. In the present study, it is hypothesized that SC can produce secretory materials, like saliva in mammals, to sterilize, to neutralize, and to digest the cystic fluid. First of all, the cyst fluid was examined for the antimicrobial property by bacteria killing assay (BKA) using E. coli, and it resulted that the cyst fluid showed relatively weak bactericidal effect. The cyst fluid was also examined for antioxidant property by DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2-azinobis-3-ethybenzthiazdine-6-sulphonic acid) assay, and resulted that the cyst fluid showed strong antioxidant effect comparable to ascorbic acid. The antioxidant materials of cyst fluid were found both in cationic and anionic groups separated by ion exchange column using SP and Q beads, and they were also heat resistant. The strong antioxidant property seems to be originated from the abundant carotenoid materials in the cyst fluid of SC. However, the strong antioxidant materials of the cyst fluid may play a role for the essential antioxidant in the digestive tract of SC. Furthermore, the cyst fluid of SC was examined using SNU-1 cancer cells for its anticancer property. The flow cytometry analysis showed that both of cationic and anionic materials purified from the cyst fluid showed strong anticancer effect on SNU-1 cells. Taken together, the cyst fluid of SC showed weak antimicrobial, but strong antioxidant property and marked anticancer properties in vitro experiment. Therefore, it is presumed that SCs can neutralize the various free radicals of seawater and also protect themselves from carcinogenic impacts in aggressive marine environment by various bioactve substances of their cyst fluid.

Insects constitute the largest and most diverse group of animals on Earth. They also serve as the hosts or nutrient sources for an immense assemblage of parasites, pathogens, and predators, ranging from bacteria and fungi to plant and animals. More than 700 known fungal species from 100 genera have adopted an entomopathogenic lifestyle. These fungi are widely distributed, occurring in aquatic, forest, and agricultural habitats, and are often used as active ingredients in microbial insect pest control agents. Their mode of action against insects involves the attachment of conidia to the insect cuticle, followed by germination, cuticle penetration, and internal dissemination throughout the insect. During this process, secreted enzymes, protein toxins, and secondary metabolites can be used by the fungus to overcome the host immune system, modify host behavior, and defend host resources against competing pathogens and saprophytes. In some cases, the host insect relationship has been found to be associated with bioactive fungal metabolites. These metabolites exhibit a wide variety of insecticidal, antibacterial, antifungal, anticancer, antioxidant, and antiviral activities. Using molecular techniques and phylogenetic analyses, both the asexual (anamorphic) stages and sexual (teleomorphic) stages of entomopathogenic fungi have recently been shown as linked together. Therefore, entomopathogenic fungi, especially in complex with the host insect, might be a promising source of bioactive molecules of pharmaceutical and various industrial interests. Here, we evaluated the antimicrobial activity of entomopathogenic fungi metabolites against plant pathogenic bacteria and fungi for the use in agriculture. The radicals scavenging activity and the anticancer activity were also evaluated for pharmaceutical interests.

쌀 도정시 발생하는 미강의 부가가치를 높이기 위하여,미강내 생리기능성 물질를 고농도로 농축하기 위한 전처리방법 및 농축조건을 확립하고자 하였다. 미강으로부터 추출한 조미강유중 지방산을 분리하기 위하여 산 또는 효소촉매를 이용하여 지방산 에틸 에스터를 제조하였다. 산 촉매 에스터 반응시 tocopherol류의 함량은 크게 영향을 받지 않았으나, tocotrienol류의 함량은 크게 감소하였고, 특히 γ-tocotrienol은 52% 감소하였다. 효소 촉매 에스터반응시 tocopherol류와 tocotrienol류 모두 크게 변화가 없었으므로 효소촉매를 이용한 에틸 에스터 반응이 생리활성물질농축에는 더 적합하였다. 효소 촉매 반응을 이용하여 생산한 미강유 에틸 에스터를 초임계 이산화탄소를 이용하여분리하고, 생리활성물질을 농축하였다. 온도 조건은 45oC,50oC, 55oC, 압력조건은 9.62MPa, 10.34MPa, 11.03MPa이었다. Tocols는 소량이지만, 1-5번 추출 분획에서 모두검출되었으며, 5번 분획에 가까워질수록 더 많은 양의tocols이 추출되었다. γ-Oryzanol도 매우 소량이기는 하나 1-5번 분획에서 추출되었다. Policosanol과 phytosterol은 1-5번 분획에서 전혀 추출되지 않았다. 특히 초임계 이산화탄소의 밀도와 지방산 에스터와 함께 추출되는 tocols 및 γ-oryzanol의 양은 높은 상관관계를 가지고 있었다(R2tocols=0.9306, R2oryzanol=0.7934). Tocols와 phytosterol은 55oC,9.62MPa에서 농축시 농축률이 가장 높았으며, γ-oryzanol과policosanol은 각 조건별로 농축률에 변화가 없었다. 또한초임계 이산화탄소의 밀도가 클수록 농축속도는 매우 빨랐으나 농축물질의 선택성은 낮았다. 따라서 생리활성물질을농축하는데에는 상대적으로 낮은 밀도의 초임계 이산화탄소를 적용하는 것이 더 효과적이었으며, 이러한 결과는 건강기능식품등에 이용하는 생리활성물질의 농축 및 정제 공정에 효율적으로 적용될 수 있을 것이다.

The content of bioactive substances and antioxidative activity in conventionally grown brown rice (CGBR) and organically grown brown rice (OGBR) were compared. Minerals (mg/100 g) such as magnesium (OGBR, 168.59±2.62; CGBR, 121.43±2.22), copper (OGBR, 0.50±0.06; CGBR, 0.41±0.05), and manganese (OGBR, 4.70±0.04; CGBR, 2.49±0.02) were higher in OGBR than in CGBR (p<0.05). In addition, levels of (μg/100 g) vitamins B2 (OGBR, 27.22±2.56; CGBR, 22.12±2.24) and B6 (OGBR, 46.32±2.66; CGBR, 39.91±3.32) were higher in OGBR than in CGBR (p<0.05). The contents (mg/100 g) of β-sitosterol (OGBR, 27.40±2.79; CGBR, 24.75±1.06), total phenolic (OGBR, 6.72±0.02; CGBR, 6.64±0.02), and ferulic acid (OGBR, 1.75±0.45; CGBR, 1.11±0.14) as well as the antioxidative activity (OGBR, 53.09±1.90%; CGBR, 48.29±3.38%) evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging assay were higher in OGBR than in CGBR, although no significant differences between samples were observed. In comparison to the control group, brown rice samples significantly inhibited cholesteryl ester hydroperoxide formation in rat plasma subjected to copper ion-induced lipid peroxidation. The inhibitory effect of OGBR was higher than that of CGBR. These results indicate that OGBR showed higher levels of bioactive substances and enhanced antioxidative activity than CGBR, although the differences were not statistically significant.