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- 저자 : 조문제 ( Moonjae Cho ) (검색결과 3 건)
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Hepatoprotective effect of Hippocampus abdominalis hydrolysate
손모아 ( Moa Son ),문준영 ( Jun Young Moon ),박상규 ( Sanggyu Park ),조문제 ( Moonjae Cho ) 한국응용생명화학회 2016 Journal of Applied Biological Chemistry (J. Appl. Vol.59 No.3
Recently, liver damage contributes to big percentage of the morbidity and mortality rates worldwide. Excessive intake of alcohol is one of the major causes of liver injury. When liver injury is repeated and becomes chronic, it leads to development of fibrosis and cirrhosis. In the liver, TGF-β is a profibrogenic cytokine, which participates in various critical events cause liver fibrosis. Seahorse (Hippocampus abdominalis) is a common traditional Chinese medicine and has been widely used for centuries. Seahorse has been known to have a variety of bioactivities, such as anti-oxidant, anti-fatigue, and anti-tumor. Peptide is one of the main compounds of seahorse. In this study, we isolated enzymatic hydrolysate from seahorse H. abdominalis by alcalase hydrolysis and investigated the effect of the hydrolysate on liver injury. In the present in vitro studies, the hydrolysate increases cell viability of Chang cells and protects Huh7 cells from ethanol toxicity. In addition, the hydrolysate inhibits TGF-β-induced responses. In vivo studies show that the pretreatment of hydrolysate reduces alcohol-induced increases of serum Glutamic oxaloacetic acid transaminase and Glutamic pyruvate transaminase activities and increases liver weight and body weight. These results suggest that seahorse may have a hepatoprotective effect.
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Xanthomonas campest Pv. glycines 8rd 에서 분리한 Glycinecin A gene promoter 연구
조문제 제주대학교 생명과학연구소 1999 제주생명과학연구 Vol.2 No.-
Xanthomonas campestris pv. glycines 8ra produces bacteriocin, glycinecin, which inhibits the growth of bacteria specifically belong to Xanthomonas spp.. Xanthomonas spp. includes many important plant pathogenic bacteria such as X. c. pv. vesicatoria which causes the bacterial leaf spot in pepper and tomato and X. oryzae pv. oryzae which causes bacterial leaf blight in rice. Glycinecin has many of good properties as a biological control agent. Since glycinecin inhibit the growth of phytopathogenic Xanthomonas sp, it can be used to control disease without any harm to beneficial bacteria for plant mostly belonged to Pseudomonas sp. In addition to that it is safe for user and environment. However, it needs some modification to develop glycinecin as a effective biological control agent. The cosmid clones involved in the production of glycinecin were isolated from the genomic library of X. c. pv. glycines 8ra by Ahn et. al., 1996. The DNA region for bacteriocin production was localized and its DNA sequences for this region had been decided. This region carries 2 putative open reading frames(ORFs) and these 2 ORFs are relevant for the activity of glycinecin in E. coli To study the regulation of glycinecin genes and environmental signals involved in the glycinecin production, promoter analysis using β-galactosidase and β-glucuronidase as a reporter was performed. X. c. pv. glycines 8ra harboring each of ORF1-uidA and ORF2-uidA fusions showed the growth phase-dependent expression of glycinecin genes. The activity of glucuronidase of ORF1-uidA and ORF2-uidA started to increase from the early stationary phase and reach the maximum at the late stationary phase.
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유기인계 및 카바메이트계 농약의 고감수성 아세틸콜린에스테라이즈의 대량생산
김영미,조문제,김소미 한국응용생명화학회 2003 Applied Biological Chemistry (Appl Biol Chem) Vol.46 No.4
본 실험에서는 acetylcholinesterase(AChE, EC 3.1.1.7)를 이용한 간이 잔류농약 검사법에 필요한, 유기인계 및 카바메이트계 살충제에 대한 감수성이 증가된 AChE(MAChE)를 baculovirus를 이용하여 대량으로 생산하는 시스템을 구축하고 생산된 효소의 특성을 관찰하였다.한라산에서 채취한 초파리에서 AChE의 cDNA를 합성한 수 PCR을 이용하여 AChE의 lipid anchor 부분을 제거하고 site directed mutagenesis에 의해 E107Y, F368L, L408F의 염기서열을 변화시켜 재조합된 MAChE cDNA를 합성하였고 baculovirus vector에 삽입하여 대량생산을 시도하였다. 대량 증식에 필요한 조건으로 감염횟수가 네 번일 때, 그리고 세포수가 2×10^(6) cell/㎖일 때 세포의 증식과 효소의 활성이 극대화됨을 알 수 있었다. His tag을 붙여 Ni-NTA affinity column을 이용하여 MAChE를 정제하였으며, 정제된 효소는 실험조건하에서는 pH(3-10)와 온도(20-50℃)의 변화에 영향을 받지 않았다. 농약 추출액으로 methanol을 사용했을 때가 ethanol을 사용할 때 보다 효과적임을 알 수 있었다. 대표적인 유기인계와 카바메이트계 농약에 대한 저해율을 조사한 결과 재조합된 MAChE는 대만의 집파리 및 변형되지 않은 AChE에 비하여 전반적으로 농약에 대한 감수성이 높은 것으로 나타났다. For the simple rapid bioassay of organophosphorus and carbamate pesticide residues, a mass-production system of acetycholinesterase (AChE, EC 3.1.1.7, MAChE) using baculovirus and insect cell culture was constructed. The cDNA for AChE was synthesized from Drosophila melanogaster in Halla Mountain, the lipid anchor tail was removed by PCR and was used for the site-directed mutagenesis of three amino acid residues (E107Y, F368L, L408F). The mutated cDNA was inserted into the baculovirus vector and expressed in insect cells. Maximum cell growth and enzyme activity were reached when the cells (2×10^(6) cell/㎖) were infected four times at four-day-intervals. His-tag containing MAChE was prufied using Ni-NTA column and used for characterization. The activity was maintained under various pHs (3-10) and temperatures (20-50℃) under experimental conditions. As an extractionn solution for pesticides, methanol is more effective than ethanol. Against major organophosphate and carbamate pesticides, the MAChE showed better sensitivity than AChE and AChE from housefly (Taiwan).
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