인삼 사포닌의 생화학적 연구 ( Ⅸ ) 인삼 사포닌의 임계 미셀 농도와 지질 분산 및 효소 활성에 미치는 영향

주충노,이상직 ( Chung No Joo,Sang Jik Lee ) 생화학분자생물학회 1977 BMB Reports Vol.10 No.2

The critical micellar concentration (CMC) of ginseng saponin was determined by osmotic pressure and electric conductivity measurements and it was found that the CMC of the saponin was about 2.0,%. When the saponin was added to chicken intestinal lumen fluid, it was realized that the fluid. was dispersed effecively, When the saponin was present together with cholesterol in the mixture, it was found that the CMC of the saponin was lowered greatly down to as small as 0.1%. Again the CMC of Na-cholate (5mM) was found lowered down below 1.0mM in the presence of 0.1% saponin. Observation of the effect of saponin on several reactions catalyzed by such enzymes as mitochondrial dehydrogenases, alcohol and aldehyde dehydrogenases, isocitrate dehydrogenases and transaminases conducted previously in this laboratory supported further the conception that surface activities of the saponin might play a significant role on the enzyme catalyzed reactions.

Biochemical Studies on Ginseng Saponins (IX) Determination of Critical Micellar Concentration of the Saponin of Korean Ginseng Roots and its Effect on Lipid Dispersion and Enzyme Reactions

주충노,이상직,Joo, Chung-No,Lee, Sang-Jik 생화학분자생물학회 1977 한국생화학회지 Vol.10 No.2

한국 인삼 뿌리(금산삼, 5년근)의 한 성분인 인삼 사포닌의 여러가지 조건하에서의 임계미셀농도(critical micellar concentration, CMC)를 측정하고 인삼 사포닌의 지질 분산 효과를 관찰하여 다음과 같은 결론을 얻었다. 1) 순수한 인삼 사포닌 용액의 CMC는 약 2%였으나 인삼 사포닌이 cholesterol과 공존할때의 인삼 사포닌의 CMC는 0.1%로 저하하였다. 2) Na-cholate가 cholestrol과 공존시의 CMC는 약 5mM였으나 이 혼합액에 인삼 사포닌을 0.1%이 되도록 가했을때 Na-cholate의 CMC는 약 1mM로 저하 되었다. 3) 또한 닭의 소장의 장강액에 인삼 사포닌을 가하면 (in vitro) 장강지질은 미셀조로 분산됨이 관찰되었다. 이와같은 실험결과로 미루어보아 인삼 사포닌은 생리적으로 중요한 담즙산이나 인지질에 의한 지질분산을 크게 도울 것으로 예측된다. 4) 여러 가지 효소(mitochondrial dehydrogenase들, alcohol dehydrogenase, aldehyde dehydrogenase, isocitrate dehydrogenase($NADP^+$), transaminase 등) 반응에 미치는 인삼 사포닌의 영향을 관찰한 연구 결과는 인삼 사포닌이 상기한 모든 효소반응에 공통적인 효과를 나타내는 것으로 풀이되며 어떤 특이한 효소에 특이적으로 작용하는 것으로는 해석하기 어려운 것으로 미루어 보아 인삼 사포닌의 표면활성이 효소반응에 크게 영향을 미칠것으로 생각된다. The critical micellar concentration (CMC) of ginseng saponin was determined by osmotic pressure and electric conductivity measurements and it was found that the CMC of the saponin was about 2.0%. When the saponin was added to chicken intestinal lumen fluid, it was realized that the fluid. was dispersed effectively. When the saponin was present together with cholesterol in the mixture, it was found that the CMC of the saponin was lowered greatly down to as small as 0.1%. Again the CMC of Na-cholate (5mM) was found lowered down below 1.0mM in the presence of 0.11% saponin. Observation of the effect of saponin on several reactions catalyzed by such enzymes as mitochondrial dehydrogenases, alcohol and aldehyde dehydrogenases, isocitrate dehydrogenases and transaminases conducted previously in this laboratory supported further the conception that surface activities of the saponin might playa significant role on the enzyme catalyzed reactions.

인삼 Saponin 류의 생화학적 연구 ( 1 ) 인삼 saponin 의 몇 가지 탈수소효소 작용에 미치는 영향

주충노,유학수,이상직,이효숙 ( Chung No Joo,Rim Soon Choi,Sang Jik Lee,Sung Hee Cho,Myung Hee Son ) 생화학분자생물학회 1973 BMB Reports Vol.6 No.3

The effect of the saponin of Panax giseng C. A. Meyer on bovine liver alcohol dehydrogenase and chick`s cardiac, major pectoral muscle and serum lactate dehydrogenases was observed and the following results were obtained. 1). Bovine liver alcohol dehydrogenase was found to be activated in the presence of th.e ginseng saponin and its maximum activity was observed at the concentration of 0.142% of the saponin in the reaction mixture. 2). Chick`s major pectoral muscle lactate dehydrogenase was activated significantly in the presence of the saponin. It was found that when the concentration of the saponin reached 0.166 in the reaction mixture, it showed maximum velocity of pyruvate reduction. When the concentration of saponin was 0.17-0.35%, the lactate oxidation occurred rapidly. 3). The ginseng saponin did not affect noticeably the activities of neither chick`s liver and cardiac nor serum lactate dehydrogenases, but large amounts of the saponin(more than 0.1-0.2% in the reaction mixture) inhibited both of them.

인삼 Saponin의 생화학적 연구(IV) 말 및 닭의 간 Alcohol Dehydrogenase에 미치는 영향

주충노,윤병희,이상직,한정호,Joo, Chung-No,Yoon, Byung-Hee,Lee, Sang-Jik,Han, Jung-Ho 생화학분자생물학회 1974 한국생화학회지 Vol.7 No.3

한국산 인삼(Panax ginseng C. A. Meyer)의 한 성분군인 saponin 류의 말 및 닭의 간 alcohol dehydrogenase(ADH)에 미치는 영향을 분광학적으로 관찰함으로써 다음과 같은 결과를 얻었다. 1) 말과 닭의 간 ADH의 $K_m$ 값은 각각 $8{\times}10^{-4}M$ 및 $5.1{\times}10^{-4}M$이다. 2) ADH, ethanol의 농도를 일정하게 유지하고 saponin의 농도를 증가시키면 saponin 농도의 증가에 따라 ADH의 활성이 점차척으로 증가하나, 반응액에서의 saponin 류의 농도가 어느 이상(말은 $0.53{\times}l0^{-3}%$ 닭은 $0.27{\times}10^{-3}%$) 이 되면 ADH 활성은 오히려 감소되었다. 3) 반응액에서의 saponin 류의 농도가 $1.33{\times}10^{-5}{\sim}6.6{\times}10^{-5}%$ 일때의 말의 간 ADH의 반응 속도와 ethanol 농도와의 관계는 sigrnoid 형의 곡선으로 표시되었다. 4) 인삼 saponin 류의 농도 뿐 아니라, ethanol 농도에 따라서도 크게 다르며 반응액에서의 인삼 saponin 류의 농도가 $1.33{\times}10^{-5}%$이고 ethanol의 농도가 2~4mM 범위일 때 말의 간 ADH는 크게 활성화됨을 관찰하였다. The effect of ginseng saponins on equine and chicken hepatic alcohol dehydrogenases was investigated spectrophotometric ally and the following results were obtained. 1) The Km values of equine and chicken hepatic ADH were found to be $8{\times}10^{-4}M$ and $5.1{\times}10^{-4}M$ respectively. 2) When the concentration of ginseng saponins of ADH reaction mixture in the presence of known amounts of ADH and ethanol, the activity of ADH increased gradually as the concentration of the saponins increased. It was found, however, that the activity of the hepatic ADH decreased when the concentration of the saponins was over $0.53{\times}10^{-3}%$ (horse) and $0.27{\times}10^{-3}%$ (chicken) respectively. 3) The velocity-concentration curves of the ADH reactions in vitro in the presence of the saponins ranging from $1.33{\times}10^{-5}%$ were found to be sigmoid. 4) It was realized that the effect of the saponins on ADH activity was dependent not only upon its concentration but also upon the concentration of substrate and that the optimal concentrations of saponins and ethanol for the maximum activity of horse ADH were $1.33{\times}10^{-5}%$ and 2~4 mM respectively.

인삼 Saponin 의 생화학적 연구 ( Ⅳ ) 말 및 닭의 간 Alcohol Dehydrogenase 에 미치는 영향

주충로,윤병희,이상직,한정호 ( Chung No Joo,Byung Hee Yoon,Sang Jik Lee,Jung Ho Han ) 생화학분자생물학회 1974 BMB Reports Vol.7 No.3

The effect of ginseng saponins on equine and chicken hepatic alcohol dehydrogenases was investigated spectrophotometrically and the following results were obtained. 1) The Km values of equine and chicken hepatic ADH were found to be 8 × 10^(-4)M and 5.1 × 10^(-4)M respectively. 2) When the concentration of ginseng saponins of ADH reaction mixture in the presence of known amounts of ADH and ethanol, the activity of ADH increased gradually as the concentration of the saponins increased. It was found, however, that the activity of the hepatic ADH decreased when the concentration of the saponins was over 0.53 × 10^(-3) % (horse) and 0.27 × 10^(-3) % (chicken) respectively. 3) The velocity-concentration curves of the ADH reactions in vitro in the presence of the saponins ranging from 1.33 × 10^(-5) % were found to be sigmoid. 4) It was realized that the effect of the saponins on ADH activity was dependent not only upon its concentration but also upon the concentration of substrate and that the optimal concentrations of saponins and ethanol for the maximum activity of horse ADH were 1.33 × 10^(-5) % and 2∼4 mM respectively.

인삼 Saponin 류의 생화학적 연구(I) -인삼 saponin의 몇 가지 탈수소효소 작용에 미치는 영향

주충노,유학수,이상직,이효숙,Joo, Chung-No,Yoo, Rak-Soo,Lee, Sang-Jik,Lee, Ryo-Sook 생화학분자생물학회 1973 한국생화학회지 Vol.6 No.3

Panax ginseng C. A. Meyer의 saponin류의 몇 가지 탈수소효소계에 미치는 영향을 관찰하고 다음과 같은 결과를 얻었다. 1) 쇠 간의 알코올 탈수소효소는 인상 saponin 존재하에서 활성화되고, 반응액의 saponin 농도가 0.142% 일때에 최고 활성을 보였다. 2) 닭의 대흉근 젖산 탈수소효소는 saponin 존재하에서 크게 활성화되고 반응액에서의 saponin 농도가 0.166% 일때에 최고 활성을 보였다. 3) 간, 삼장 및 혈청 젖산 탈수소효소는 인상 saponin에 의해 크게 활성화되지 않으며 다량의 인삼 saponin 존재하에서는 오히려 억제된다. The effect of the saponin of Panax giseng C. A. Meyer on bovine liver alcohol dehydrogenase and chick's cardiac, major pectoral muscle and serum lactate dehydrogenases was observed and the following results were obtained. 1). Bovine liver alcohol dehydrogenase was found to be activated in the presence of the ginseng saponin and its maximum activity was observed at the concentration of 0.142% of the saponin in the reaction mixture. 2). Chick's major pectoral muscle lactate dehydrogenase was activated significantly in the presence of the saponin. It was found that when the concentration of the saponin reached 0.166% in the reaction mixture, it showed maximum velocity of pyruvate reduction. When the concentration of saponin was 0.17-0.35%, the lactate oxidation occurred rapidly. 3). The ginseng saponin did not affect noticeably the activities of neither chick's liver and cardiac nor serum lactate dehydrogenases, but large amounts of the saponin (more than 0.1-0.2% in the reaction mixture) inhibited both of them.

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 Saponin 의 생화학적 연구 ( 3 ) 닭의 간 미토콘드리아의 몇가지 탈수소 효소에 미치는 영향

주충노,최임순,정노팔,이상직,김옥희 ( Chung No Joo,Rim Soon Choe,Ro Pal Chung,Sang Jik Lee,Ok Hee Kim ) 생화학분자생물학회 1974 BMB Reports Vol.7 No.1

The effects of ginseng saponins on chick`s hepatic mitochondrial dehyrogenases were oberved using the dye 2, 6-dichlorophenolindophenol (DICPIP) as oxidant. The reduction of DICPIP was measured spectrophotometrically and the following results were obtained. 1. Although chick liver contains much NAD after homogenation, it is well known that NAD is destroyed fairly rapidly by NADase in the liver and virtually no NAD is present by the time the experiment being carried out. It was observed in this experiment that the addition of NAD to the reaction mixture containing mitochondrial preparation restored the action of the dehydrogenases and their activities were raised as the concentration of saponin in the reaction mixture increased and the maximum activity was observed when 1.2% saponin solution was added (the saponin concentration of the reaction mixture was 0.17%, approx. 1.7 mM), indicating that the ginseng saponin might activate NAD-dependent dehydrogenases. 2,. When α-ketoglutarate, one of the TCA cycle intermediates and/or L-glutamate was added to the above reaction mixture, the reduction of DICPIP was greatly increased in the presence of the ginseng saponin suggesting that the saponin activated α-ketoglutarate dehydrogenase and/or L-glutamate dehydrogenase. 3. It was also observed that the oxidation of succinate by either hepatic or cardiac mitochondrial preparation of chicken was increased significantly as the concentration of saponin in the reaction mixture increased, showing that the saponin activated flavin-dependent dehydrogenase.

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.

인삼 Saponin의 생화학적 연구 (III) 닭의 간 미토콘드리아의 몇가지 탈수소 효소에 미치는 영향

주충노,최임순,정노팔,이상직,김옥희,Joo, Chung-No,Choe, Rim-Soon,Chung, Ro-Pal,Lee, Sang-Jik,Kim, Ok-Hee 생화학분자생물학회 1974 한국생화학회지 Vol.7 No.1

인삼 saponin이 닭의 간 미토콘드리아 탈수소효소에 미치는 영향을 DICPIP (2, 6-dichlorophenolindophenol)의 환원을 분광학적으로 측정함으로써 조사하여 다음과 같은 결과를 얻었다. 1. 간미콘드리아 조제물에 $NAD^+$를 가하면 미토콘드리아의 탈수소효소가 재활성화하며, 인삼 saponin의 농도가 증가함에 따라 그 활성화도가 커지고 1.2%의 saponin 용액을 가했을 때(반응액의 saponin 농도는 0.17%, 약1.7 mM) 최고활성을 보였다. 이것은 saponin이 NAD-linked dehydrogenere의 활성화를 의미하는 것으로 생각된다. 2. TCA 회로의 중요유기산인 $\alpha$-ketoglutarase나 또는 중요아미노산인 L-glutamate를 간 미토콘드리아 조제물에 가하면 saponin의 농도가 증가함에 따라 유기산의 산화가 크게 촉진됨이 관찰되었으며, saponin이 $\alpha$-ketoglutarate dehydrogenase나 Lglutamate dehydrogenase를 활성화함이 예측된다. 3. TCA 회로 효소인 succinate dehydrogenase도 saponin에 의하여 크게 활성화됨이 예측된다. The effects of ginseng saponins on chick's hepatic mitochondrial dehyrogenases were oberved using the dye 2, 6-dichlorophenolindophenol (DICPIP) as oxidant. The reduction of DICPIP was measured spectrophotometrically and the following results were obtained. 1. Although chick liver contains much NAD after homogenation, it is well known that NAD is destroyed fairly rapidly by NADase in the liver and virtually no NAD is present by the time the experiment being carried out. It was observed in this experiment that the addition of NAD to the reaction mixture containing mitochondrial preparation restored the action of the dehydrogenases and their activities were raised as the concentration of saponin in the reaction mixture increased and the maximum activity was observed when 1.2% saponin solution was added (the saponin concentration of the reaction mixture was 0.17%, approx. 1.7 mM), indicating that the ginseng saponin might activate NAD-dependent dehydrogenases. 2. When $\alpha$-ketoglutarate, one of the TCA cycle intermediates and/or L-glutamate was added to the above reaction mixture, the reduction of DICPIP was greatly increased in the presence of the ginseng saponin suggesting that the saponin activated $\alpha$-ketoglutarate dehydrogenase and/or L-glutamate dehydrogenase. 3. It was also observed that the oxidation of succinate by either hepatic or cardiac mitochondrial preparation of chicken was increased significantly as the concentration of saponin in the reaction mixture increased, showing that the saponin activated flavin-dependent dehydrogenase.