Aldehyde Dehydrogenase의 반응 메카니즘에 관한 연구 -Dehydrogenase 활성의 속도결정 단계의 추구-

주충노,김두식,구현서,김주휘,Joo, Chung-No,Kim, Doo-Shik,Koo, Hyon-Seo,Kim, Joo-Hwee 생화학분자생물학회 1992 한국생화학회지 Vol.25 No.6

소의 간 aldehyde dehydrogenase(ALDH)와 사람 적혈구 ALDH에 미치는 금속이온의 영향과 화학변형 실험을 행하고 보고된 ALDH의 반응 메카니즘에 관한 실험결과들을 종합하여 광범위한 특이성을 가진 일반 ALDH의 dehydrogenase 활성의 속도제한 단계가 Enzyme-NADH complex로부터의 NADH의 해리단계임을 알 수 있었다. The effect of several ions and chemical modification experiments using bovine liver cytosolic aldehyde dehydrogenase (ALDH) and human erythrocyte ALDH have been carried out to understand the rate-limiting process of dehydrogenase activity. From the previously reported experimental results and this study suggested that the releasing step of NADH from Enzyme-NADH complex might be a rate-limiting process.

알데히드 탈수소효소 촉매반응의 특성

주충노,김두식,황우익 ( Chung No Joo,Doo Sik Kim,Woo Ik Hwang ) 생화학분자생물학회 1991 BMB Reports Vol.24 No.5

The effect of several modifiers including disulfiram and phenylmethanesulfonylfluoride and metal ions on the dehydrogenase and esterase activities of several aldehyde dehydrogenase (ALDH) isozymes isolated from rat liver and brain, bovine brain and baker`s yeast was investigated to see whether or not the both reactions share a cmomon active site. Several lines of experimental evidence suggested that ALDH might have a common active site for both reactions, but with different microenvironmental situations for dehydrogenase and esterase reactions.

Aldehyde Dehydrogenase 의 반응 메카니즘에 관한 연구 - Dehydrogenase 활성의 속도결정 단계의 추구

주충노,김두식,구현서,김주희 ( Chung No Joo,Doo Shik Kim,Hyon Seo Koo,Joo Hwee Kim ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.6

The effect of several ions and chemical modification experiments using bovine liver cytosolic aldehyde dehydrogenase (ALDH) and human erythrocyte ALDH have been carried out to understand the rate-limiting process of dehydrogenase activity. From the previously reported experimental results and this study suggested that the releasing step of NADH from Enzyme-NADH complex might be a rate-limiting process.

Catalytic Properties of Aldehyde Dehydrogenase

주충노,김두식,황우익,Joo, Chung-No,Kim, Doo-Sik,Hwang, Woo-Ik 생화학분자생물학회 1991 한국생화학회지 Vol.24 No.5

The effect of several modifiers including disulfiram and phenylmethanesulfonylfluoride and metal ions on the dehydrogenase and esterase activities of several aldehyde dehydrogenase (ALDH) isozymes isolated from rat liver and brain, bovine brain and baker's yeast was investigated to see whether or not the both reactions share a cmomon active site. Several lines of experimental evidence suggested that ALDH might have a common active site for both reactions, but with different microenvironmental situations for dehydrogenase and esterase reactions. 쥐의 간과 뇌, 소의 뇌, 효모균에서 추출 정제한 몇 가지 aldehyde dehydrogenase(ALDH) isozyme에 미치는 disulfiram, phenylmethanesulfonyl-fluoride 등과 같은 modifier와 금속이온 등의 영향을 관찰하고 ALDH의 dehydrogenase 반응과 esterase 반응의 활성부위의 동일성 여부를 추구한 결과 ALDH의 dehydrogenase 반응과 esterase 반응의 활성부위는 같으나 주변의 미세환경에 차이가 있는 것으로 생각된다.

인삼 사포닌의 생화학적 연구 ( ⅩⅠ ) 인삼 사포닌이 Aldehyde Dehydrogenase 에 미치는 영향

주충노,구자돈,김두식,이상직 ( Chung No Joo,Ja Don Koo,Doo Sik Kim,Sang Jik Lee ) 생화학분자생물학회 1977 BMB Reports Vol.10 No.2

It was attempted in this study to observe the effect of ginseng saponins, one of the major components of Korean ginseng root, on rat hepatic aldehyde dehydrogenase (ALDH, EC 1. 2. 1.3) to understand some physiological role of the saponin on the alcohol metabolism in the body and the following results were obtained: 1) It was realized that mitochondria) ALDH was stimulated by the saponin, the concentration of which were around 1 × 10^(-8)%-1 × 10^(-6)% in the assay mixture but inhibited when the saponin concentration was over 1 × 10^(-4)%. 2) Km of mitochondria) ALDH for acetaldehyde reached the lowest value at 1 × 10^(-7)% of the saponin suggesting that the saponin stimulation of the ALDH might be brought by Km lowering for acetaldehyde of ALDH. Cytosolic ALDH was also found stimulated by the saponin. 3) Alcohol dehydrogenase (ADH, EC 1. 1. 1. 1. ) stimulation by moderate amounts of the saponin was confirmed in the present experiment using radioactive ethanol-^(14)C and it has been suggested that the moderate amount of the saponin accelerated ethanol oxidation to acetate by stimulating both ADH and ALDH.

Biochemical Studies on Ginseng Saponins(X) The Effect of Ginseng Saponins on Lipid Metabolism

주충노,이희봉,김두식,Joo, Chung-No,Lee, Hee-Bong,Kim, Doo-Sik 생화학분자생물학회 1977 한국생화학회지 Vol.10 No.2

한국산 인삼(Panax ginseng C.A. Meyer) 뿌리의 주요 성분 중의 하나인 인삼 사포닌류가 지방산의 산화와 합성에 미치는 영향(in vitro)과 아울러, 동위원소 $^{14}C$으로 표지된 acetate-1, 2-$^{14}C$을 쥐에 투여하여 지질대사에 미치는 영향(in vitro)을 관찰하고 다음과 같은 결과를 얻었다. 1. 인삼 사포닌은 쥐의 간 미토콘드리아에 의한 지방산 산화와 시토졸에 의한 지방 합성을 다같이 촉진하였다. 2. 쥐에게 인삼 사포닌을 투여하고 4시간 후 acetate-1, 2-$^{14}C$을 투여하여 적당 시간후(5분~240분)의 간장, 뇌, 지방 조직 및 혈청의 지질 방사능의 분포를 조사하였다. 3. 혈청을 제외한 다른 상기조직에서의 지질 방사능은 실험군에서는 30분에서 60분사이에 큰 방사성이 관찰되었고, 대조군의 경우는 2 시간 후에나 큰 방사성이 나타나는 것으로 미루어 보아 인삼 사포닌이 상기 조직에서 지방 합성을 크게 촉진한 것으로 생각된다. 4. 본 연구의 실험 조건하에서 측정된 간 조직의 콜레스테롤과 지방산의 생물학적 반감기는 대조군의 경우, 각각 30분과 70분인데 비하여 실험군은 19분과 44분이었다. 이와같은 실험 결과는 인삼 사포닌이 콜레스테롤과 지방산의 대사(분해와 합성)를 촉진하는 것으로 해석된다. 5. 뇌에서의 콜레스테롤과 지방산의 최고 방사능은 실험군의 경우 동위원소 주입 후 각각 15분과 30분이고 대조군의 경우는 각각 216분과 156분이었으며, 인삼 사포닌이 뇌의 지질대사를 크게 촉진하는 것으로 관찰되었다. 6. 지방조직에서는 실험군의 최고 방사능이 동위원소 주입 후 61분이었고 대조군은 124분이었으며, 인삼 사포닌의 동위원소의 지방산으로의 도입을 크게 촉진하였다. 7. 혈청 지질의 최고 방사능은 실험군의 경우 동위원소 주입 후 79분, 대조군은 216분이었으며, 인삼사포닌은 지질대사뿐 아니라 지질의 이동에도 크게 영향을 미치는 것으로 예측된다. It was attempted to observe the effects of ginseng saponins, one of the major components of Korean ginseng roots, on fatty acid oxidation and synthesis in vitro as well as on lipid metabolism in vivo and the following results were obtained. 1. It was found that the ginseng saponins stimulated significantly both fatty acid oxidation by rat hepatic mitochondrial preparation and synthesis by the cytosol fraction. 2. Distribution of radioactivities of lipids in liver, brain, adipose tissue and blood serum of albino rats administered with the ginseng saponins prior to acetate -1, 2-$^{14}C$ injection intraperitoneally have been investigated. 3. The high radioactivities of lipid in the above tissues of test group except blood serum appeared at about 30 min. to 60 min. after acetate injection while those of control group was observed at about two hours after suggesting that the ginseng saponins stimulated the biosynthesis of lipids in the above tissues. 4. Under the experimental conditions in this study, it was realized that biological half lives of hepatic cholesterol and fatty acid of test rat were found to be 19min. and 44min. while those of the corresponding lipids of control rat were 30min. and 70min. respectively suggesting that the ginseng saponins might stimulate the metabolism, both biosynthesis and degradation of cholesterol and fatty acid of the liver of this animal. 5. In the brain, the highest specific radioactivities of cholesterol and fatty acid of test group were observed at 15min. and 30min. after acetate-1, 2-$^{14}C$ injection while those of the corresponding lipids of control group were found at 216min. and 156min. after the injection respectively suggesting that the ginseng saponins stimulated greatly the metabolism of brain lipids. 6. In the adipose tissue, the highest specific radioactivity of fatty acid of test group was observed at 61min. after acetate-1, 2-$^{14}C$ injection while that of the control group appeared at 124 min. after the injection suggesting that the ginseng saponin stimulated Carbon-14 incorporation into fatty acid in this tissue. 7. It appeared that the high radioactivity of blood serum lipid of control group was observed at 216min. after acetate-1, 2-$^{14}C$ injection but the radioactivity peak of test group was observed at 79 minutes after the injection of 1, 2-$^{14}C$-acetate signifying that the ginseng saponin affected not only lipid metabolism but also lipid transport in the animal body.

Biochemical Studies of Ginseng Saponins(XI) The Effect of Ginseng Saponins on Aldehyde Dehydrogenase

주충노,구자돈,김두식,이상직,Joo, Chung-No,Koo, Ja-Don,Kim, Doo-Sik,Lee, Sang-Jik 생화학분자생물학회 1977 한국생화학회지 Vol.10 No.2

이상이 되면 ALDH반응은 오히려 억제되었다. 2) ALDH의 acetaldehyde에 대한 Km은 $1{\times}10^{-7}%$의 인삼 사포닌 존재시 최소의 값 (1.67mM)이였으며 (Control의 Km은 2.74mM) 인삼 사포닌에 의한 ALDH반응의 촉진은 ALDH의 acetaldehyde에 대한 Km의 저하에 기인하는 것으로 추측된다. 시토졸 분획의 ALDH반응도 인삼 사포닌에 의하여 촉진되었다. 3) 인삼 사포닌에 의한 alcohol dehydrogenase (ADH, EC 1. 1. 1. 1) 반응의 촉진현상은 ethanol-1-$^{14}C$을 사용한 본 연구에서도 확인되었으며 적당량의 인삼 사포닌은 ADH 반응과 ALDH반응을 다같이 촉진함으로서 체내에서의 ethanol 산화와 유독한 acetalde-hyde의 제거를 원만히 진행하는 것으로 생각된다. It was attempted in this study to observe the effect of ginseng saponins, one of the major components of Korean ginseng root, on rat hepatic aldehyde dehydrogenase (ALDH, EC 1. 2. 1. 3) to understand some physiological role of the saponin on the alcohol metabolism in the body and the following results were obtained: 1) It was realized that mitochondrial ALDH was stimulated by the saponin, the concentration of which were around $1{\times}10^{-8}%-1{\times}10^{-6}%$ in the assay mixture but inhibited when the saponin concentration was over $1{\times}10^{-4}%$. 2) Km of mitochondrial ALDH for acetaldehyde reached the lowest value at $1{\times}10^{-7}%$ of the saponin suggesting that the saponin stimulation of the ALDH might be brought by Km lowering for acetaldehyde of ALDH. Cytosolic ALDH was also found stimulated by the saponin. 3) Alcohol dehydrogenase (ADH, EC 1. 1. 1. 1.) stimulation by moderate amounts of the saponin was confirmed in the present experiment using radioactive ethanol-1-1-$^{14}C$ and it has been suggested that the moderate amount of the saponin accelerated ethanol oxidation to acetate by stimulating both ADH and ALDH.

인삼 사포닌의 생화학적 연구 ( Ⅹ ) 한국산 인삼 사포닌의 지질대사에 미치는 영향

주충노,이희봉,김두식 ( Chung No Joo,Hee Bong Lee,Doo Sik Kim ) 생화학분자생물학회 1977 BMB Reports Vol.10 No.2

It was attempted to observe the effects of ginseng saponins, one of the major components of Korean ginseng roots, on fatty acid oxidation and synthesis in vitro as well as on lipid metabolism in vivo and the following results were obtained. 1. It was found that the ginseng saponins stimulated significantly both fatty acid oxidation by rat hepatic mitochondrial preparation and synthesis by the cytosol fraction. 2. Distribution of radioactivities of lipids in liver, brain, adipose tissue and blood serum of albino rats administered with the ginseng saponins prior to acetate -1, 2-^(14)C injection intraperitoneally have been investigated. 3. The high radioactivities of lipid in the above tissues of test group except blood serum appeared at about 30 min. to 60 min. after acetate injection while those of control group was observed at about two hours after suggesting that the ginseng saponins stimulated the biosynthesis of lipids in the above tissues. 4. Under the experimental conditions in this study, it was realized that biological half lives of hepatic cholesterol and fatty acid of test rat were found to be 19min. and 44min. while those of the corresponding lipids of control rat were 30min. and 70min. respectively suggesting that the ginseng saponins might stimulate the metabolism, both biosynthesis and degradation of cholesterol and fatty acid of the liver of this animal. 5. In the brain, the highest specific radioactivities of cholesterol and fatty acid of test group were observed at 15min. and 30min. after acetate-1, 2-^(14)C injection while those of the corresponding lipids of control group were found at 216min. and 156min. after the injection respectively suggesting that the ginseng saponins stimulated greatly the metabolism of brain lipids. 6. In the adipose tissue, the highest specific radioactivity of fatty acid of test group was observed at 61min. after acetate -1, 2-^(14)C injection while that of the control group appeared at 124 min. after the injection suggesting that the ginseng saponin stimulated Carbon-14 incorporation into fatty acid in this tissue. 7. It appeared that the high radioactivity of blood serum lipid of control group was observed at 216min. after acetate-1, 2-^(14)C injection but the radioactivity peak of test group was observed at 79 minutes after the injection of 1, 2-^(14)C-acetate signifying that the ginseng saponin affected not only lipid metabolism but also lipid transport in the animal body.

인삼 사포닌이 지용성 비타민(A와 E)의 흡수에 미치는 영향

朱忠魯,金斗植,全英運 연세대학교 자연과학연구소 1978 學術論文集 Vol.1 No.-

본 연구에서는 人蔘 Saonin 이 不水溶性 Vitamin인 Vitamin A와 Vitamin E의 吸收에 미치는 영향을 쥐(albino rat, 100~120g ♂)를 使用하여 觀察하고 다음과 같은 結果를 얻었다. 1) Vit. A 를 人蔘 Saponin 과 함께 Stomach tubing法으로 投與한후 쥐의 血淸을 조사한 結果 試驗群의 Vit.A의 含量은 對照群보다 2倍나 된다. 그러나 3時間後부터는 過量의 Vitamin含量이 減少되기 시작하였다. 試驗群의 肝의 Vitamin A含量도 對照群의 1.5倍였으며 3時間後에는 正常 Vitamin量으로 回復되었다. 2) Vit.E의 경우도 Vitamin E를 投與하고 40分後에 血淸과 肝에서의 Vit. E含量을 調査한 結果 試驗群이 對照群에 比하여 1.4~1.5倍였다. 3) 위와같은 實驗結果는 人蔘 Saponin이 Vit. A와 Vit. E의 吸收를 促進한다는 것을 意味하는 것이다. 4) 한편 試驗群의 腎藏에서의 Vitamin含量이 對照群에 比하여 훨씬 높은 것은 人蔘 Saponin이 不水溶性 Vitamin의 吸收뿐 아니라, 移動 및 排泄에도 크게 영향을 미칠것으로 예상된다. It was attempted in this experiment to observe the effect of ginseng saponin on the absorption of water insoluble Vitamins such as Vitamin A and Vitamin E using albino rat (150-200g,♂) and the following result were obtained. 1) When the Vitamin A and ginseng saponin were fed by stomach tubing, Vit. A concentration of blood serum of test group was found as much as twice that of control, but the Vitamin A content started to decrease 3 hours after. The Vitamin A content of the liver of test animal was also found about 1.5 times that of control and its value returned to normal value three hours later. 2) The Vitamin E contents of the blood and lover of test animals were found as much as 1.4-1.5 times that of control 40 minutes after feeding. 3)The above results suggested tha saponin stimulated the absortion of water insolubl Vitamin A and E 4) On the other hand, the contents of the above Vitamins in the kidney of test animals were significantly higher suggesting that the ginseng saponin also affects the transport of the above water insoluble Vitamins.