• Title/Summary/Keyword: Metabolizing Enzyme

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Effect of Maesil (Prunus mume) Juice on the Alcohol Metabolizing Enzyme Activities (매실즙이 알코올대사 효소활성에 미치는 영향)

  • Hwang, Ja-Young;Ham, Jae-Woong;Nam, Sung-Hee
    • Korean Journal of Food Science and Technology
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    • v.36 no.2
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    • pp.329-332
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    • 2004
  • Changes in activities of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) in vitro were examined by measuring maximum absorbances of ADH and ALDH at 340 nm to determine influence of Maesil (Prunus mume) on alcohol metabolism. Facilitating rates of ADH activity were 137.92, 131.58, 152,96, 218.70, 111.76, and 144.27% in Maesil juice, 5, 10, and 15% GMT, and 0.5 and 1.0% aspartic acid, respectively, ALDH activity increased in the order of Maesil juice > ALDH > GMT > aspartic acid, and facilitating rate of ALDH activity in Maesil juice was the highest at 976.44%. These results indicate alcohol metabolizing activity can be enhanced by Maesil juice.

The Effects of Houttuynia cordata on the Hepatic Bromobenzene Metabolizing Enzyme System in Rats and Isolation of Phenolic Compounds (흰쥐의 브로모벤젠대사계에 미치는 어성초의 영향과 페놀성 화합물의 분리)

  • Hur, Jong-Moon;Park, Ju-Gwon;Park, Sung-Jong;Lee, Jong-Ho;Sung, Nak-Ju;Choi, Myeong-Rak;Song, Sang-Ho;Kim, Moon-Sung;Choi, Jong-Won;Park, Jong-Cheol
    • Korean Journal of Pharmacognosy
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    • v.31 no.2
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    • pp.228-234
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    • 2000
  • Effects of Houttuynia cordata on the level of lipid peroxide and the enzyme activities of the liver were investigated in bromobenzene-induced rats. Lipid peroxide content in liver was increased by bromobenzene. It was decreased when the methanol extract from the aerial parts of H. cordata was treated to the rat. The methanol extract reduced the activities of aminopyrine N-demethylase and aniline hydroxylase that increased by bromobenzene, however did not affect glutathione S-transferase activity. The methanol extract recovered the activity of epoxide hydrolase activity that decreased significantly by bromobenzene. We suggest that under our experimental conditions the extract might play an important play in the prevention of hepatotoxicity by reduction of aminopyrine N-demethylase and aniline hydroxylase activities as well as enhancement of epoxide hydrolase activity. Six phenolic compounds have been isolated from H. cordata and identified by means of spectral analysis as protocatechuic acid, quercetin, apigenin, afzelin, hyperoside and quercitrin.

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Overexpression of the get Gene Encoding 4-α-Glucanotransferase of a Hyperthermophilic Archaeon, Thermococcus litoralis (초호열성 고세균 Thermococcus litoralis로부터 4-α-glucanotransferase의 대량밭현)

  • Jeon, Beong-Sam;Park, Jeong-Won;Shin, Gab-Gyun;Kim, Beom-Kyu;Kim, Hee-Kyu;Song, Jae-Young;Cho, Young-Su;Cha, Jae-Young
    • Journal of Life Science
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    • v.14 no.3
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    • pp.435-440
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    • 2004
  • The gene encoding a extremely thermostable 4-$\alpha$-glucanotransferase from a hyperthermophilic archaeon, Thermococcus litoralis, was cloned, sequenced and expressed in Escherichia coli. The amino acid sequence of the enzyme was distantly related to other functionally-related ones, such as D-enzymes. The enzyme is of industrial interest because of a novel activity of producing cycloamylose and is also important for fundamental studies of protein, sugar-metabolizing enzymes. In this paper, the overexpression of 4-$\alpha$-glucanotransferase in E. coli was carried out expression vector system with lac and T7 promoters. The enzyme was successfully overexpressed, and purified by the heat treatment of a cell-free extract, successive Butyl-Toyopearl and Mono Q chromatographies. The purified recombinant enzyme showed the same specific activity and the same mobility in SDS-PAGE as natural enzyme.

Changes of Hepatic Cyclohexane Metabolizing Enzyme Activities and Its Metabolites in Serum and Urine after Cyclohexane Treatment

  • Kim Ji-Yeon;Jeon Tae-Won;Lee SangHee;Chung Chinkap;Joh Hyun-Sung;Lee Sang-Il;Yoon Chong-Guk
    • Biomedical Science Letters
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    • v.11 no.4
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    • pp.509-515
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    • 2005
  • This study was conducted to determine the kinetics of cyclohexane metabolites (the biomarker on cyclohexane exposure), the changes of hepatic cyclohexane metabolizing enzyme activities and the metabolites of cyclohexane in urine or serum. The rats were sacrificed at 2, 4, 8, 12 and 24 hr after administration of one dose of cyclohexane (1.56 g/kg body weight, i.p.). The metabolites of cyclohexane in urine were identified as cyclohexanol, cyclohexanone, trans-l,2-cyclohexanediol and 1,4-cyclohexanediol with cyclohexane metabolite being 124.00, 0.78, 23.28 and 2.75 (g/g of creatinine, $1\times10^{-3}$). Most of the cyclohexanol and trans-l,2-cyclohexanediol were determined to be in the form of $\beta-glucuronide$ conjugates, whereas cyclohexanone and 1 ,4-cyclohexanediol were found as free forms. In toxicokinetics of serum cyclohexane metabolites, cyclohexanol showed a rapid increase, reaching the plateau at 4 hr, after this time rapidly decreased throughout 24 hr. Changes of cyclohexanone also showed the similar pattern with cyclohexanol except somewhat lower concentration. Trans-l,2-cyclohexanediol, however, showed a gradual increase until 12 hr with the continued same levels throughout 24 hr. On the other hand, 1,4-cyclohexanediol was detected as trace levels at 4 and 12 hr, respectively. The administration of cyclohexane led to a significant increase of hepatic aniline hydroxylase activity from 2 to 8 hr. The activity of hepatic alcohol dehydrogenase showed a significant increase at 4 hr and then were recovered to the level of the control at 24 hr. On the other hand, there were no differences in liver weightlbody weight between the control and cyclohexane-treated animals. However, there were the changes of aniline hydroxylase and alcohol dehydrogenase activities on time-dependent pattern after cyclohexane treatment, which influence on the degree of cyclohexane metabolites both in blood and urine. These results suggest that differential determination of cyclohexane metabolites in urine and serum may be able to be as a biomarker of cyclohexane-exposure in the body. But in this fields further study is needed.

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Responses in Hepatic Xenobiotic Metabolizing Enzymes and Sex Hormones of Yellowfin Goby Acanthogobius flavimanus in Nakdong Estuary (낙동강 하구에서 채집한 문절망둑 Acanthogobius flavimanus의 간장 약물대사효소계와 성호르몬 농도)

  • Lee, Ji-Seon;Jeong, Jee-Hyun;Han, Chang-Hee;Shim, Won-Joon;Jeon, Joong-Kyun
    • Korean Journal of Environmental Biology
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    • v.26 no.2
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    • pp.87-93
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    • 2008
  • To assess effects of contaminants on fish in Nakdong river, feral yellowfin goby Acanthogobius flavimanus were caugt in two different sites and its hepatic monooxygenase enzyme, including cytochrome P450 (CYP), NADPH-cytochrome P450 reductase (P450R), NADH-cytochrome b5 reductase (b5R), ethoxyresorufin deethylase (EROD), glutathione S-transferase (GST) were quantitatively determined. Gonadosomatic index (GSI), hepatosomatic index (HSI) and three sex steroid hormone (17$\beta$-estradiol, E2; testosterone, T; 11-ketotestosterone, 11-KT) levels of the fish were also investigated. HSI of fish from polluted site (site 1) were significantly higher than that of unpolluted site (site 2), but GSI levels were significantly lower in polluted site. No significant differences in plasma 11-KT and T levels were observed in two sites surveyed. E2 level was, however, significantly (p<0.05) higher in female fish from site 1 than site 2. In addition, hepatic EROD activity and CYP level of site 1 fish were lower than those of site 2 fish, whereas relatively high levels of P450R, b5R and GST activities were found in site 1. The results imply that yellowfin goby, especially female fish in Nakdong river estuary are affected from contaminants disrupting sex steroid hormone system.

Comparative liver drug metabolizing enzymes activities between Korean native cattle and swine (한우와 돼지에서 간의 약물 대사효소의 활성 비교)

  • Lee, Gwan-bok;Yun, Hyo-in;Park, Seung-chun;Kim, Min-kyu;Lee, Rae-kyung;Cho, Joon-hyung;Lee, Dong-woo
    • Korean Journal of Veterinary Research
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    • v.38 no.1
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    • pp.17-28
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    • 1998
  • Drug-metabolizing activities of Korean native cattle and swine were investigated from viewpoints of the cytochrome P-450's level, their dependent mixed function oxidase activities, the reactive oxygen species formation and cytosolic enzyme acitivities from each liver homogenates. Level of cytochrome P-450 in the liver microsome of Korean native cattle was $0.28{\pm}0.05nmole/mg$ and that in pigs $0.35{\pm}0.03nmole/mg$. Level of cytochrome $b_5$ of Korean native cattle was $0.24{\pm}0.06nmole/mg$, and that of pigs $0.2{\pm}0.05nmole/mg$, showing no difference between two species. NADPH P-450 reductase were higher in Korean native cattle ($58.3{\pm}5.3nmole/mg/min$) than in pigs ($29.9{\pm}3.8nmole/mg/min$)(p<0.01). The activities of cytochrome P-450 dependent monooxygenases such as ethoxyresorufin O-deethylase (cattle, $96.5{\pm}12.5nmole/mg/min$ ; pigs, $13.6{\pm}2.1nmole/mg/min$), N-benzphetamine N-demethylase (cattle, $5.23{\pm}0.82nmole/mg/min$ ; pigs, $0.76{\pm}0.3nmole/mg/min$) and aniline hydroxylase (cattle, $0.95{\pm}0.1nmole/mg/min$ ; pigs, $0.33{\pm}0.08nmole/mg/min$) were much higher in Korean native cattle than in swine(p<0.01). However, the activity of testosterone $7{\alpha}$-hydroxylase was higher in swine ($90.4{\pm}1.2nmole/mg/min$) than cattle (cattle, $32.8{\pm}1.2nmole/mg/min$). Interestingly, testosterone $16{\alpha}$-hydroxylase, a marker enzyme for P-450 IIA was not detected in both animal species. These results suggest that Korean native cattle and pigs have high contents of P-450 IA1 and P-450 IIIA. Total sulfhydryl compound (cattle, $10.3{\pm}1.1nmole/mg$ ; Pigs, $14.5{\pm}1.8nmole/mg$) and glutathione related enzymes except glutathione reductase (cattle, $38.1{\pm}7.9nmole/mg/min$; swine, $22{\pm}3.6nmole/mg/min$) showed higher levels in swine than in Korean native cattle. Superoxide dismutase (cattle, $7.64{\pm}0.84nmole/mg/min$ ; pigs, $4.47{\pm}0.94nmole/mg/min$) and catalase (cattle, $30.4{\pm}3.7nmole/mg/min$ ; pigs, $17.2{\pm}1.8nmole/mg/min$) were remarkably higher in Korean native cattle than in swine (p<0.05).

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Effects of Yuldahanso-tang and Chungsimyonja-tang on Cytochrome P450 Activities (열다한소탕과 청심연자탕의 Cytochrome P450 활성 연구)

  • Jin, Seong-Eun;Ha, Hye-Kyung;Shin, Hyeun-Kyoo
    • Journal of Sasang Constitutional Medicine
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    • v.24 no.4
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    • pp.84-91
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    • 2012
  • Objectives : The purpose of this study is to investigate the inhibitory or inductive potentials of Yuldahanso-tang (YDT) and Chungsimyonja-tang (CST), herbal formulas for Taeeumin, on cytochrome P450 (CYP450) drug metabolizing enzyme. The mechanisms for the herbal formula-drug interaction has not been well reported in spite of the chance for co-administration with conventional drugs. Methods : To evaluate the interaction potential of YDT-drug or CST-drug, the fluorescence-based enzyme assays on CYP450 isozymes including CYP3A4, CYP2C19, CYP2D6 and CYP2E1 were established in vitro. The inhibitory effects of herbal formulas were characterized with $IC_{50}$ values. Results : YDT showed inhibitory effects on CYP2D6 and CYP2E1-mediated metabolism, while it exhibited week inhibition on CYP3A4 and CYP2C19 relatively. CST exerted relatively weak inhibitory effects on the four CYP450 isozymes compared to that of YDT. Conclusions : These results suggest that the herbal formula-drug interaction could be occur when YDT are co-administered with drugs mediated by CYP2D6 or CYP2E1.

Effect of Ginseng on the Hepatic Alcohol Metabolizing Enzyme System Activity in Chronic Alcohol-Treated Mouse (만성(慢性) Acohol 섭취(攝取) mouse에서 alcohol 대사(代謝) 효소활성(酵素活性)에 미치는 인삼(人蔘)의 영향(影響))

  • Choi, Chong-Won;Lee, Sang-Ill;Huh, Keun
    • The Korean Journal of Pharmacology
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    • v.20 no.1 s.34
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    • pp.13-21
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    • 1984
  • The present study was undertaken to investigate an effect of ginseng butanol fraction(total saponin) on the hepatic ethanol metabolism, we used experimental animals for the subject of study. When, in case of ADH and MEOS, ginseng butanol fraction was added, enzyme activity was increased in a small dose, and on the contrary, in a large dose, showed inhibitory effect. in catalase, the activity showed no significant effect by adding ginseng butanol fraction. In the light of kinetic aspect, when, in reaction mixture, ethanol and ginseng butanol fraction were concurrently added and reacted, Km value of ADH and MEOS was decreased. After pretreated with ginseng butanol fraction and inducement of acute toxic state by ethanol, the activities of ADH and MEOS were increased to an extent of about 25% compared to controls. But catalase activity was not significantly affected. In case that ginseng butanol fraction was given to mice fed with 5% ethanol instead of water for 60 days, the activities of ADH and MEOS were increased about 20% to 50% compared to ethanol-treated group. On the contrary, catalase activity was not affected. But blood concentrations of ethanol were decreased due to ginseng butanol fraction treatment. All these observations suggested that reduction of ethanol blood concentration should be dependent upon increased activities of ADH and MEOS. Thereby it suggests the recovery from alcohol intoxication can be prompted by treatment with ginseng.

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Cloning of Phospholipase D from Grape Berry and Its Expression under Heat Acclimation

  • Wan, Si-Bao;Wang, Wei;Wen, Peng-Fei;Chen, Jian-Ye;Kong, Wei-Fu;Pan, Qiu-Hong;Zhan, Ji-Cheng;Tian, Li;Liu, Hong-Tao;Huang, Wei-Dong
    • BMB Reports
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    • v.40 no.4
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    • pp.595-603
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    • 2007
  • To investigate whether phospholipase D (PLD, EC 3.1.4.4) plays a role in adaptive response of post-harvest fruit to environment, a PLD gene was firstly cloned from grape berry (Vitis Vinifera L. cv. Chardonnay) using RT-PCR and 3'- and 5'-RACE. The deduced amino acid sequence (809 residues) showed 84.7% identity with that of PLD from Ricinus communis. The secondary structures of this protein showed the characteristic C2 domain and two active sites of a phospholipid-metabolizing enzyme. The PLD activity and its expression in response to heat acclimation were then assayed. The results indicated PLD was significantly activated at enzyme activity, as well as accumulation of PLD mRNA and synthesis of new PLD protein during the early of heat acclimation, primary suggesting that the grape berry PLD may be involved in the heat response in post-harvest grape berry. This work offers an important basis for further investigating the mechanism of post-harvest fruit adaptation to environmental stresses.

The Effects of Dietary Proteins on Hepatic Histological Changes and Membrane Stability in Chemically Induced Rat Hepatocarcinogenesis (화학적으로 유도한 랫드 간세포 암화과정에서 단백질 식이가 간의 조직학적 변화와 생체막 안정도에 미치는 영향)

  • 박경애;김현덕;최혜미
    • Journal of Nutrition and Health
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    • v.34 no.8
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    • pp.833-842
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    • 2001
  • The purpose of this study is to determine the effect of dietary proteins and fats on the hepatic histological changes, membrane stability, and drug-metabolizing enzyme activities during chemically induced rat hepatocarcinogenesis. Weanling Sprague-Dawley rats were fed the diet containing 20% casein or soy protein isolate and 15% perilla or corn oil for 10 weeks. Hepatocarcinogensis was initiated with diethylnitrosamine(DEN), and the rats were fed diets containing 0.02% 2-acetylaminofluorene(AAF) followed by 0.05% phenobarbital (PB). The scores of histological changes were decreased in treated rats fed soy protein diet compared to those find casein diet. Liver weights were significantly increased by AAF and PB treatment in rats fed casein diets in both oil groups. Glucose 6-phosphatase(G6Pase) activities, an index of membrane stability, were significantly reduced by AAF and PB treatment in rats find casein diets, and were lower in casein diet compared to soy protein diet groups. Especially, the activities were the highest in the rats fed soy protein-perilla oil diet. Lipid peroxide values also were increased by AAF and PB treatment in rats fed casein diet. Aniline hydroxylase activities were not influenced by protein and fat sources. Glutathione-dependent enzyme activities were increased by AAF and PB treatment. Linoleic and arachidonic acid content were increased in rats fed corn oil diet, and linolenic and eicosapentaenoic acid contents were increased in rats fed perilla oil diet. Our results suggest that soy protein isolate inhibit the abnormal histological changes in liver, possibly by maintaining the membrane stability during chemically induced rat hepatocarcinogenesis. Soy protein may be protective against the hepatocarcinogenesis induced by chemical carcinogen.

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