면역시스템을 통한 세포수명연장 천연물질 스크린

유보경(Bo-Kyung Yoo),권기상(Kisang Kwon),고영화(Young Hwa Ko),김홍근(Hong Geun Kim),이석현(Seokhyun Lee),박관호(Kwan-Ho Park),최지영(Ji-Young Choi),권오유(O-Yu Kwon) 한국생명과학회 2016 생명과학회지 Vol.26 No.3

한약진흥재단(NIKOM)의 640-천연물 라이브러리를 PC12세포에 10 ㎍/ml/day로 처리하였다. 면역강화에 관여하는 ERAP1 (Endoplasmic Reticulum AminoPeptidase 1)과 수명연장에 관여하는 FOXO1 (FOXO-family transcription factor) 유전자발현을 RT-PCR로 확인하였다. ERAP1 유전자를 1.5-2배까지 상승시킨 것은 22개, 2배 이상 발현시킨 것은 14개였다. FOXO1 유전자를 1.5-2배까지 상승시킨 것은 총 24개, 2배 이상 발현시킨 것은 4개였다. 동시에 ERAP1과 FOXO1 유전자를 2배 이상 상승시키는 것은 족도리풀(Asiasari radix)에서 분리된 (±)-Car-3-ene-2,5-dione, 두꺼비(Bufonis venenum)에서 분리된 Cinobufagin, 대국(Euphorbiae pekinensis)에서 분리된 Corilagin, 현호색(Corydalis tuber)에서 분리된 Corydaline 4종이다. 이들 4종의 천연물은 초파리를 이용한 개체의 수명연장실험에 사용될 것이다. We have screened four natural products against 640 single compounds, which shows more two folds gene expression for both endoplasmic reticulum aminopeptidase 1 (ERAP1) and FOXO-family transcription factor (FOXO1). The results were as follows. (±)-Car-3-ene-2,5-dione from Asarum sieboldii Miq. is C10H12O₂ molecular formula and the 164 kDa molecular weight. Cinobufagin from Bufonis Venennum is C26H34O₆ molecular formula and 442 kDa molecular weight. So far reported main biological function is Na⁺/K⁺-ATPase inhibition. Corilagin from Euphorbia pekinensis is C27H22O18 molecular formula and 634 kDa molecular weight. Carbonic anhydrase inhibition is well known its biological function. Corydaline from Corydalis turtschaninovii is C22H27NO₄ molecular formula and 369 kDa molecular weight. The main biological function is acetylcholinesterase inhibition. In the short future, four types of natural products will be used in longevity experiments with insects. The results may give one of the clues for studying new drug development candidates of the longevity.

장수풍뎅이(Allomyrina dichotoma)에 Oryctes rhinoceros nudivirus 감염에 의해서 유전자 발현이 조절되는 5개의 유전자

유보경(Bo-Kyung Yoo),권기상(Kisang Kwon),고영화(Young Hwa Ko),이은령(Eun Ryeong Lee),최지영(Ji-Young Choi),권오유(O-Yu Kwon) 한국생명과학회 2016 생명과학회지 Vol.26 No.11

최근에 장수풍뎅이(Allomyrina dichotoma)는 관상용으로 인기가 높으며, 애벌레는 건강보조식품으로 주목을 받고 있다. 장수풍뎅이의 대량사육 시 발생하는 질병의 원인이 Oryctes rhinoceros nudivirus (OrNV)인 것이 2015년에 처음 보고되었다. 그러나 아직 정확한 진단, 발병기전과 치료방법을 찾지 못하고 있으며 곤충 사육농가에 매년 심각한 경제적 손실을 입고 있다. 본 연구는 장수풍뎅이의 대량사육 시 발생하는 OrNV질병의 조기진단과 치료에 실마리를 제공하기 위하여 OrNV에 감염된 장수풍뎅이의 장(intestine)에서 유전자발현이 조절되는 Klf15, ERAP2, Snrnp200, mbnl2a, MIMI_L93를 보고한다. Korean rhinoceros beetles (Allomyrina dichotoma), which can be found in broad-leaved forests in mountainous habitats and lives for around one year in wild. This beetle is currently popular as a pet and traditionally regarded as a medicine for liver-related diseases in Korea. It is reported that the economic losses in the mass-rearing facilities by virus infection have been increased since the 2010s in Korea. The causing virus for the A. dichotoma was firstly reported as an Oryctes rhinoceros nudivirus (OrNV) in 2015. We, here, observes that serious morphological changes in the intestinal tube from the OrNV-infected beetles, and report five genes, which are regulated by OrNV infection in the intestine; Krueppel-like factor 15 (Klf15), Endoplasmic reticulum aminopeptidase 2 (ERAP2), U5 small nuclear ribonucleoprotein 200 kDa helicase (Snrnp200), Muscleblind-like protein 2a (mbnl2a), and MIMI_L93. The results may provide a clue to the early diagnosis and disease treatment during the mass-rearing facilities of the A. dichotoma.

한약재 추출물에 대한 전혈응집억제 효능 검색

전원경 ( Won Kyung Jeon ),유보경 ( Bo Kyung Yoo ),김영은 ( Yeong Eun Kim ),박선옥 ( Sun Ok Park ),박선민 ( Sun Min Park ),고병섭 ( Byoung Seob Ko ) 한국응용생명화학회 2007 Applied Biological Chemistry (Appl Biol Chem) Vol.50 No.4

장수풍뎅이(Korean Rhinoceros Beettle, Allomyrina dichotoma)에 Oryctes rhinoceros nudivirus 감염 특이적으로 발현하는 유전자 동정

권기상(Kisang Kwon),유보경(Bo-Kyung Yoo),서현우(Hyun-Woo Suh),고영화(Young Hwa Ko),김홍근(Hong Geun Kim),이석현(Seokhyun Lee),박관호(Kwan-Ho Park),최지영(Ji-Young Choi),권오유(O-Yu Kwon) 한국생명과학회 2015 생명과학회지 Vol.25 No.8

장수풍뎅이(Korean Rhinoceros Beettle, Allomyrina dichotoma)는 관상용으로 인기가 높으며, 애벌레는 식품으로 주목을 받고 있다. 장수풍뎅이의 대량사육 시 발생하는 질병의 원인이 최근에 virus (Oryctes rhinoceros nudivirus)인 것이 보고되었다. 그러나 아직 정확한 발병기전과 치료방법을 알지 못하여 사육농가에 심각한 경제적 손실을 주고 있다. 본 연구에서는 virus에 감염된 장수풍뎅이의 fat body와 장에서 심각한 조직변화가 관찰되었다. 이 때에 유전자발현이 상승하는 것으로 BTF3H4-like, SPS-like, COPB1, T-CP, HSP70를 보고한다. 이 결과는 장수풍뎅이의 대량사육 시 발생하는 질병의 조기진단과 치료에 실마리를 제공할 것이다. 그리고 안정적인 생산성 향상을 통해서 소득증대 효과 및 균일화된 대량사육을 통해서 식품원료로 인정받을 수 있을 것이다. The Korean rhinoceros beetle (Allomyrina dichotoma) is popular as a pet and as a food ingredient, and it is commercially distributed in Korea. It is also traditionally regarded as a medicine for liver-related diseases. Recently, the Oryctes rhinoceros nudivirus was introduced from Southeast Asia. This virus is reported as a disease factor for A. dichotoma in mass-rearing facilities, and economic losses due to this viral infection have been increasing in Korea since the 2010s. In this study, we observed serious structural changes in the fat body and the intestine of virus-infected beetles. We report five genes that are up-regulated by the viral infection in the intestine: BTF3H4-like (transcription factor BTF3 homolog 4-like), SPS-like (serine proteinase stubble-like), COPB1 (coatomer protein complex, subunit beta 1), T-CP (T-complex 1 subunit gamma), and HSP70 HSP70 (heat shock protein 70). The results may provide a clue for the early diagnosis and disease-treatment that occurs in mass-rearing facilities. The improvement of stable productivity will increase the farmers’ income, and quality control of beetle-breeding will help industries to utilize this beetle as a promising food ingredient.

Isolation and Characterization of Six Microorganisms from the Digestive Tract of the Cricket Gryllus bimaculatus

Kisang Kwon(권기상),Eun Ryeong Lee(이은령),Bo-Kyung Yoo(유보경),Young Hwa Ko(고영화),Hyojung Shin(신효정),Ji-Young Choi(최지영),O-Yu Kwon(권오유) 한국생명과학회 2017 생명과학회지 Vol.27 No.9

우리는 처음으로 쌍별귀뚜라미 소화기관에서 6종류의 미생물을 분리하고 특성을 규명하였다. 이들은 16S rDNA을 기준으로 분류한 결과 4종류(Staphylococcus, Bacillus, Citrobacter, Proteus)에 속하였다. 분리된 6종류의 미생물은 공통적으로 ampicillin에 저항성을 보이지만 kanamycin 저항성은 보이지 않았다. 이들을 Gram염색하여 미생물의 형태적 특징을 확인하였다. Gram-positive한 rod-shaped GL2와 round-shaped GL4는 다른 분리 균 보다 많은 양의 세포외분비물을 만들었다, 이들을 MALDI-TOF-MS spectral analysis결과 87-kDa collagenase, 56-kDa & 200-kDa hypothetical protein이였다. 새롭게 분리된 6종류의 미생물은 귀뚜라미의 생리에 미치는 영향과 이들의 생물공학적 혹은 해충 방제에 이용될 수 있는 연구가 기대된다. We describe the isolation and characterization of six different intestinal microorganisms from the digestive tract of the cricket Gryllus bimaculatus. Based on 16S rRNA gene sequences, we obtained six isolates belonging to four different genera: Staphylococcus, Bacillus, Citrobacter, and Proteus. All the isolates were resistant to ampicillin. Ampicillin is an irreversible inhibitor of the enzymeetranspeptidase, which is needed to make bacterial cell walls. None of the isolates were resistant to kanamycin, which binds to the 30S subunit of the bacterial ribosome and then inhibits total protein synthesis. Gram staining was conducted, in addition to morphological classification under a microscope. Four grampositive isolates and two gram-negative isolates were detected. The gram-positive isolates were GL1 (round shaped, 2 am in diameter), GL2 (rod shaped, 2.5 μm in length), GL3 (rod shaped, 2 μm in length), and GL4 (round shaped, 1.5 μm in diameter). The gram-negative isolates were GL5 (rod shaped, 2 μm in length) and GL6 (rod-shaped, 2.5 μm in length). Notably, two of the isolates, GL2 and GL4, secreted specific extracellular proteins. These were determined by MALDI-TOF-MS spectral analysis to be a 87 kDa collagenase, 56 kDa hypothetical protein, and 200 kDa hypothetical protein. The six isolates in this study could be used for various biotechnological applications and pest management, both in the field and in greenhouse systems. In addition, it would be interesting to determine the relationship between these isolates and their host.

천연물에 의한 초파리수명연장 효과

이정훈(Jeong Hoon Lee),권기상(Kisang Kwon),이은령(Eun Ryeong Lee),유보경(Bo-Kyung Yoo),고영화(Young Hwa Ko),최지영(Ji-Young Choi),권오유(O-Yu Kwon) 한국생명과학회 2017 생명과학회지 Vol.27 No.1

우리는 이전실험에서 배양세포를 사용하여 4종류의 천연물(Corydaline, (±)-Car-3-ene-2,5-dione, Cinobufagin, Corilagin)이 ERAP1와 FOXO1 (DFA16) 유전자발현을 2배 이상 상승시키는 것을 증명하였다. 본 실험은 1% agar, 5% sucrose, natural compound 20 μl를 넣은 먹이를 만들어 4시간 starvation후에 4시간 동안 먹였다. Cinobufagin와 Corilagin를 먹이면 대조군에 비하여 6-8일 정도 더 생존하였다. RT-PCR 실험결과 ERAP1와 FOXO1 유전자 발현을 조절하는 것이 증명되었다. 산업곤충질병 진단과 치료에 사용될 것이며, 초파리를 대신한 생쥐에서도 동일한 결과를 실험하여 수명연장을 위한 신약으로 발전시킬 것이다. A previous study demonstrated that four kinds of natural compounds, Corydaline, (±)-Car-3-ene-2,5-dione, cinobufagin, and corilagin, enhanced the gene expression of ERAP1 and FOXO1 (DFA16) more than two-fold in a cell culture system. In this study, the experimental food was made finally 30 ml in which included 1% agar, 5% sucrose, and each natural compound 20 μl. A fruit fly of Drosophila melanogaster fed a natural product for 4 hr after 4 hr starvation. Both natural compounds of Cinobufagin and Corilagin induced 6-8 days more survival comparing than it controls group. The resulting fruit flies were estimated the gene expression of ERAP1 and FOXO1 by RT-PCR that also demonstrated meaningful results with the same lifespan results. Cinobufagin from BufonisVenennumis has C26H34O₆ molecular formula and 442 kDa molecular weight. Corilagin from Euphorbiapekinensisis has C27H22O18 molecular formula and 634 kDa molecular weight. The two types of natural products screened in this study will be used in the early diagnosis and treatment of insect industry in the near future. In addition, the natural products will be used in longevity experiments in a mouse model. The results may give one of the clues for studying new drug development candidates of the longevity.