γ-P roteobacteria에 속하는 신종균 DS-01에서 생성되는 Agarase의 생화학적 특성

한원동,박권필,서민영,황영민,성치남 한국엔터테인먼트산업학회 2012 한국엔터테인먼트산업학회논문지 Vol.6 No.2

해수에서 새로이 분리된 균주 DS-01이 생성하는 agarase를 부분 정제하여 생화학적 특성을 규명하였다. Agarase의 최적 pH는 6.0, 최대반응온도는 45℃이었으며, 60℃의 열처리에 의해서도 60% 이상의 잔존활성을 보이는 열안정성 효소이었으며, Fe++와 Zn++에 의해서 90% 이상 활성이 저해되었다. 10mM 이상의 NaCl 농도에서 활성이 저해되기 시작하여 0.3M 이상에서는 활성이 60% 이상 감소되었다. 열 건조시의 안정성을 분석한 결과, 30℃에서 79%, 45℃에서도 73%의 잔존 활성도를 보여주었으며, 기질 친화도는 agar와 agarose에 대해서는 유사한 활성도를 보여 주었으나, sodium alginate는 분해하지 못하였다. 전기영동적으로 단일 band로 존재함이 확인되었다. Physicochemical property of agarase from novel marine bacterium designated strain DS-01 in the phylum Proteobacteria were studied. The maximum agarolytic activity in the reducing sugar assay was observed at 45℃ and pH 6.0. Approximately 60% residual activity was detected over all temperature(20-60℃) and pH ranges(pH 5-9) tested. The agarase activity was inhibited by Fe2+and Zn2+. This agarase showed the affinity on agar and agarose, but not on sodium alginate. Agarase from DS-01 was single protein without isozyme.

사람唾液의 Cl 血型分泌型에 關한 硏究

朴淳元,韓元東 大韓法醫學會 1979 대한법의학회지 Vol.3 No.1

Moon and Wiener had demonstrated new blood factor "Cl" with anti-Cl lectin prepared from extracts of seeds of Korean Clerodendron trichotomum THUNBERG by absorption of selected human red blood cells. After that, Moon et al. had reported that Cl type specific substance in distributed in human saliva, too. Therefore, human saliva can be classified into two type, secretor of Cl (Sc) and non-secretor of Cl (nSc). To study on the frequency of Sc and nSc, relation with ABH type secretor and hereditary pattern of Cl secretors, agglutination inhibition test of anti-Cl lectin was carried out with human saliva by standard method, and the results can be summarized as follows; 1. Cl positive type can be classified secretor and non-secretor of Cl by human saliva, but Cl negative type is reacted as only non-secretor of Cl. 2. The secretor status of Cl is quite different from ABH secretor. 3. It appears that the inheritance of the saliva type of Cl follows the Mendelian law with the non-secretor of Cl transmitted through recessive genes.

미국흰불나방(Hyphantria cunea Drury) Invertase의 특성 및 합성부위에 관한 연구

한원동,윤치영,김학열 한국곤충학회 1992 Entomological Research Vol.22 No.2

배추흰나비(Pieris rapae L.) β-N-Acetylglucosaminidase의 활성분포와 합성기관

윤치영,한원동 대전대학교 기초과학연구소 1991 自然科學 Vol.2 No.-

배추흰나비 (Pieris rapae L.)의 키틴 분해효소 중에서 3종의 β-N-acetylglucosaminidases (EⅠ, EⅡ and EⅢ)의 활성분포와 합성기관을 조사하였다. 조직배양 후 배양액 내에 존재하는 기간별 효소활성은 시간 경과에 따라 지방체에서는 큰 차이를 보이지 않은 반면, 혈림프에서는 전용중기 및 말기 재료에서 8시간 이후 뚜렷한 활성증가를 보였으며, 체벽의 경우는 전용초기의 재료에서 12시간까지 계속적인 증가를 보였다. 또한 5령 말기유충의 체벽 배양시, 20-hydroxyecdysone 처리군은 20시간까지 활성증가를 나타내었으며 비처리군은 16시간까지 증가 경향은 나타내었지만 그 이후의 양상은 현저히 달랐다. 이러한 결과들은 혈림프와 체벽내의 각 효소활성이 출현시기와 변태시의 역할이 다름을 암시할 뿐만아니라 주로 EⅠ과 EⅢ는 체벽에서, EⅡ는 혈림프에서 각각 활성을 나타내는 효소임을 보여 주었으며 또한EⅠ는 체벽에서, EⅡ는 지방체에서 각각 합성되는 것으로 추정되었다. Among chitinolytic enzymes from cabbage whitebutterfly, Pieris rapae L., distribution and synthetic organ of β-N-acetylglucosaminidases (EⅠ, EⅡ and EⅢ) were investigated. After tissue culture, enzymatic activities of each organ in the media were measured with time processing. In fat body little change was detected. On the other hand, specific increase after 8hr at early and late prepupae were detected in haemolymph, and the activity increased continuously until 12hr at early prepupae in integument. And in culturing the integument of late th instar larvae, the enzymatic activity of 20-hydroxyecdysone treated group increased significantly up to 20hr culturing while that of control group increased till 16hr but quite different pattern was detected after that time. These results showed that the enzymatic activities in haemolymph and integument differed and that they had different roles in pupal ecdysis. also these showed that EⅠand EⅢ had main activities in integument and EⅡ in haemolymph, and suggested that EⅠ was synthesized in integument and EⅡ in fat body, respectively.

이질바퀴(Periplaneta americana L.) 소화관내 invertase의 특성

이진원,심재철,조현욱,한원동 順天大學校 1997 論文集 Vol.16 No.1

이질바퀴 (Periplaneta americana)의 invertase를 분리, 분획하여 isozyme의 존재여부, 물리화학적 특성, 기관별 분후 및 변태단계에 따른 활성의 변화 등을 조사하여 다음과 같은 결과를 얻었다. P. americana의 소화관 invertase는 pH 5.5, 50℃에서 최대의 활성을 보였으며, Km 값은 ?? 의해서는 활성이 촉진되었고, ?? 의해서는 활성이 저해되었다. 열에 대한 안정성은 55℃의 열처리에서도 높은 활성을 유지하였으나, 60℃의 열처리에서는 활성이 거의 소실되었다. 변태단계별 invertase의 활성의 변화를 보면, 난협에는 invertase의 활성이 거의 없었으나, 1령유충시기부터 3령유충까지 급격한 활성의 증가현상을 보이다가 그 후로는 완만한 활성의 증강양상을 나타내었으며, 소화관에는 두 개의 isozyme(Inv-1, Inv-2)이, 말피기관과 지방체에는 Inv-2만이 존재하였고, 혈림프에는 Inv-1이 존재하였으나, 난소에는 invertase가 존재하지 않았다. invertase의 분자량은 Inv-1이 112Kd, Inv-2가 51Kd이었다. Gut invertase of Periplaneta americana was isolated and fractionated to determine the isozyme localization, physicochemical properties, organ distribution and developmental changes. There were two isozymes, Inv-1 and Inv-2, of invertase of gut from P. americana with M.W. 112Kda for Inv-1 and 51Kda for Inv-2, and their activities occurred mainly in gut, Malphigian tubule, haemolymph, and somewhat in fat body. The optimum pH of invertase of gut from P. americana was around pH 5.5 and optimal temperature was 50℃. Km value of this enzyme was ??. The activities of invertase from P. americana were activated by ?? and ??, but were inhibited by ??. This enzyme was stable for heat treatment below 55℃, but was unstable above 60℃. Activities were not appeared in egg mass, but drastically increased from the first instar nimph to the third instar nymph, and then gradually increased. Two isozymes were present in whole body and gut, but only one isozyme, Inv-2 in Malphgian tubule and fat body, and Inv-1 in haemolymph, and no one in ovary.

이질바퀴 (Periplaneta americana L.)와 독일바퀴 (Blattella germanica L.)의 invertase에 대한 생화학적 연구

한원동,주현미 한국곤충학회 2001 Entomological Research Vol.31 No.2

이질바퀴와 독일바퀴 invertase의 생화학적 특성을 조사하였다. 이질바퀴와 독일바퀴의 invertase는 각각 pH 5.0~5.2, 5.4에서 최대 활성을 보였으며, Km간은 1.43$\times$$10^{-2}$ M, 1.25$\times$$10^{-2}$ M이었다. 이질바퀴는 5$0^{\circ}C$에서 최대활성을 보이고 55$^{\circ}C$ 열처리에도 비교적 안정한 반면 독일바퀴는 35$^{\circ}C$에서 최대활성을, 5$0^{\circ}C$이상의 열처리에서 활성이 급격히 소실되었다. 이질바퀴는 $Na^{+}$, $Mn^{2+}$, $Ca^{2+}$, $K^{+}$, Fe$^{2+}$에 의해 활성이 촉진, Hg$^{2+}$에 의해 저해되었으며 독일바퀴는 Hg$^{2+}$와 Cu$^{2+}$에 의해 활성이 저해되었다. 이질바퀴에는 5개의 isozyme(Inv 1~5)이 존재하며 이 중 Inv 3는 유충기에만 나타났고, 독일바퀴에는 2개의 isozyme(inv 1, 2)이 존재하였으며, 정제된 Inv 1은 분자량이 74 Kd로 확인되었다. 한편 이질바퀴의 invertase는 독일바퀴의 invertase와 면역학적으로 서로 다른 것으로 나타났다. 이질바퀴의 invertase는 소화관, 혈림프, 지방체, 말피기써소관, 난소에도 분포함이 확인되었으며, 소화관에는 Inv 1, 2가, 지방체와 말피기씨소관에는 Inv 1, 혈림프에는 Inv 2가 각각 존재하였다. Invertases of Periplaneta americana L. and Blattella germanica L. were investigated for their physicochemical properties and isozyme localization. The optimal pH of invertase from P. americana was found to be around pH 5.0~5.2, Km value was 1.43$\times$10$^{-2}$ M, and optimal temperature was 5$0^{\circ}C$. The activities of invertase from p. ameticana were activated by Na$^{+}$, Mn$^{2+}$, $Ca^{2+}$, $K^{+}$, Fe$^{2+}$ but inhibited by Hg$^{2+}$. The optimal pH of invertase from B. germanica was found to be pH 5.4, Km value was 1.25$\times$10$^{-2}$ M, and optimal temperature was 35$^{\circ}C$. The activities were inhibited by Hg$^{2+}$ and Cu$^{2+}$. In heat stability of invertases, that of P. americana was more stable than that of B. germanica. There were five isozymes of invertase (Inv 1~5) in P. americana, and molecular weight of Inv 1 was 74 Kd. While in B. germanica, there were two isozymes (inv 1, 2) at all developmental stages. We observed that Inv 1, 2 were present in whole body and midgut, but only one isozyme, Inv 1 in fat body and malpighian tubule, and Inv 2 in haemolymph. Inv 1 and Inv 2 of P. americana appeared immunologically homozygous, but heterozygous with invertase from B. germanica.om B. germanica.

배추흰나비 (Pieris rapae L. ) 의$\beta-N-Acetylglucosaminidase$의 특성과 기능

윤치영,한원동김학열 한국통합생물학회 1993 동물학회지 Vol.36 No.1

곤충의 탈피작용에서 일어나는 키틴 분해대사를 구명하기 위한 목적으로 배추흰나비 (Pieris rupae L.)에서 3종의 서N-acetylglucosaminldase(El, Ell, Elll)를 분리, 정제하였다. Polvacrylamide gel 상에서 이들 효소의 활성은 기질과 triphenvltetrazolium chloride와의 반응 결과 생기는 발색정도로 확인하였다. 각각의 분자량은 76,000, 55,000, 35,000 da, pl 값은 모두 5.8이었으며 후p$\beta$GlcNAc와 각p$\beta$GaINAc에 대해서 다같이 기질특이성을 나타내었다. 또한 El과 Elll는 탈피 시기와 일치하는 전용 말기에 최대 활성을 나타내었으며 Ell는 용화 직후에 최대 활성을 나타내었다.

미국흰불나방(Hyphantria cunea Drury)의 Trehalas의 特性 및 合成部位에 關한 硏究

韓元東,尹治榮 順天大學校 1989 論文集 Vol.8 No.1

미국흰불나방(Hyphantria cunea Drury)의 炭水化物 加水分解酵素인 trehalase를 分離,精製하여 分離特性 및 subunit 分子量,生合成部位 등을 조사하였다. trehalase는 3개의 isozyme으로 存在하나 이들 모두가 chromatography에 의하여 단일 peak로 分離되어지는 陽 ion性의 蛋白質이었으며, 分子量 40K의 monomer로 存在하였다. 또한 trehalase의 酵素活性部位와 抗原決定基부위는 서로 다른 것으로 나타났다. trehalase의 3개 isozyme 중 消化館에만 存在하는 trehalase-3는 消化館 중에서도 中腸組織에서만 合成되며, 합성 즉시 腸內공으로 分泌되어진다. 한편 脂肪體에서는 trehalase-3가 合成되지 않는다. Trehalase of Hyphantria cunea D. were isolated and purified to determine physicochemical properties, and its synthetic portion was studied. All isozymes of trehalase were eluted single fraction after chromatography through DEAE-Sephadex A-50 column and its isozymes were determined to be monomer with M.W.40,000K dalton. According to Rocket immunoelectrophoresis, trehalase was found that active sites of enzyme does not corresponds with antigenic determinant group. Trehalase-3 which is present only in digestive tract is synthesized in the midgut tissue and then released into gut lumen, immediately. On the other hand, trehalase-3 is not synthesized in fat body.

사람 血淸의 Rat赤血球에 對한 凝集活性에 關한 硏究

韓元東,朴永學 大韓法醫學會 1985 대한법의학회지 Vol.9 No.1-2

For the purpose of individual identification, a lot of researches concerned to classification of human blood type were performed. The author experimented the agglutination activity of human sera to rat red cell, and also investigated the relation with the existing blood types to comparative possibly. The following results were obtained. 1. Human sera could be clasified into 2 groups by their agglutination activities to rat RBC: one was agglutination group, and the other was no-agglutination group. 2. Of the 131 donors, the incidence of the former (agglutination group) was 79% and the latter was 21%. 3. There was no significant relation between agglutination property of rat RBC with human sera and individual differences of rat. 4. The agglutination property of human sera to rat RBC had no relation with blood types, such as ABO, Cl, H, Vp, ab.C, CCl, and CCN etc. or serum types classified by duck, cat and bovine RBC.

Physiological Effects on Periplaneta americana of Bacillus sp. Exotoxin

임채식,한원동 한국곤충학회 2002 Entomological Research Vol.32 No.2

This study was performed to evaluate the physiological effects of Bacillus sp. on Periplaneta americana. The insecticidal exotoxin was fractionated by ion-exchange chromatography and gel filtration from Bacillus sp. cultivates. Lp2 among 4 lipoproteins showed markable increase after 24 hrs, but there was no change in glycoprotein. The enzyme activities of P. americana showed mostly decreasing pattern by treatment of exotoxin fractions. Protease activities decreased at 12 hrs after treatment in comparison with normal subjects. Amylase also showed decreasing pattern and maximal decline was observed after 12 hrs. Trehalase activities decreased at 6 hrs and 24 hrs. Invertase activities decreased from 6 hrs to 24 hrs, but recovered after 48 hrs. Amylase isozyme A1 and A2 disappeared and new isozyme appeared after 12 hrs with exotoxin treatment. Trehalase isozyme T1 and invertase I4 increased markedly after 12 hrs. However, non--speci.c esterase E1 disappeared after 24 hrs.