• Title/Summary/Keyword: enzyme induction

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Purification and In Vitro Translation of Penicillium verruculosum Cellulase mRNA

  • Kim, Jeong-Ho;Chung, Ki-Chul;Kang, Hyun-Sam;Lee, Young-Kyu
    • Journal of Microbiology and Biotechnology
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    • v.1 no.4
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    • pp.232-239
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    • 1991
  • Caboxymethyl cellulase (CMCase) I was purified from the induced culture filtrate of Penicllium verruculosum F-3 by ammonium sulfate precipitation, DEAE-Sephadex A-50 chromatography and Bio-gel P-150 filtration. The purified enzyme was assumed to be a glycoprotein consisting of 8.5% carbohydrate and having a molecular weight of 70.000 in SDS-polycrylamide gel electrophoresis (SDS-PAGE). The purified enzyme-specific anti-CMCase I IgG was obtained by rabbit immunization and protein A-sepharose CL-4B chromatography. The fungal poly($A^+$) RNA was isolated from the total RNA of the mycelium grown under cellulase induction conditions by oligo(dT)-cellulosse chromatography. The translation products in vitro were prepared by translating the isolated poly ($A^+$) RNA in rabbit reticulocyte lysate and analyzed by SDS-PAGE and fluorography. Of the translation products, CMCase I was identified by the immunoprecipitation against anti-CMCase I IgG.

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Effect of scoparone on the hepatic sulfatransferase activity in mice

  • Huh, Keun;Park, Jong-Min;Shin, Uk-Seob;Lee, Sang-Il
    • Archives of Pharmacal Research
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    • v.13 no.1
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    • pp.51-54
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    • 1990
  • Effect of scoparone (6, 7-dimethoxycoumarin) on the hepatic cytosolic sulfotransferase activity was investigated. After treatment with scoparone, hepatic cytosolic sulfotransferase activity was increased with odse and time-dependent manner as compared to control. The $V_{max}$ value (control = 1.33 n moles/mg protein/min, scoparone = 2.39n moles/mg protein/min) without affecting the $K_m$ value for p-nitrophenol was increased by the scoparone treatment. Whereas, the hepatic cytosolic sulfotransferase was not changed by the addition of scoparone in vitro, and was strongly inhibited by the addition of metabolites of scoparone. The results obtained suggest that the characteristics of increase in the enzyme activity may include induction of enzyme proteins, and may be due to the metaboltes of scoparone.

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Biochemical Properties of Haemolymph Carboxylesterase in Diapausing Pupae of Helicoverpa assulta (Guenee) (담배나방의 휴면 용 혈림프 Carboxylesterase의 생화학적 특성)

  • 김영관;이형철;박희윤;이옥경;유종명
    • Journal of the Korean Society of Tobacco Science
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    • v.20 no.1
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    • pp.71-79
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    • 1998
  • Haemolyph carboxylesterases induced in diapausing pupae of Helicoverpa assulta Guenee were investigated. Increase in the activity of the electrophoresed isozyme bands were observed during the diapausing pupae. The isozymatic composition exhibited remarkable alterations represented as disappearance and induction of some isozyme bandsp which were identified as carboxylesterase (CE) on the basis of their specificities to inhibitors. Much higher activity of the induced CE was shown in reaction with $\beta$-naphthyl acetate ($\beta$-Na) than $\alpha$-naphthyl butyrate ($\alpha$-Nb), representing the high regioselectivity to $\beta$-naphthyl group. Optimal temperature for the enzyme activity was different to the substrates used 37$^{\circ}C$ in $\beta$-Na and 4$0^{\circ}C$ in $\alpha$-Nb, respectively. However, the optimal pH for the enzyme activity was the same as 7.5 regardless of the substrates used, and relatively high thermostability of the CE was demonstrated by showing the denaturation at high temperature (50~55$^{\circ}C$).

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Wogonin, a flavone from Scutellaria radix, inhibits nitric oxide production from RAW 264.7 cells

  • Kim, Hee-Kee;Cheun, Bong-Sun;Kim, Young-Ha;Kim, Sung-Yong;Kim, Hyun-Pyo
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1998.11a
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    • pp.196-196
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    • 1998
  • Nitric oxide is involved in various physiological processes. Among isoforms of nitric oxide synthase, iNOS is partly responsible for inflammation and septic shock. During our continual search for anti-inflammatory flavonoids, we have found that flavonoids, especially flavones, possessed the inihibitory activity of NO production by iNOS from LPS-activated RAW 264.7 cell. In this study, flavonoids from Scutellaria radix were investigated for their inhibitory activity of nitric oxide production. It was found that wogonin, among tested flavonoids including baicalein, oroxylin A, skullcapflavone II, showed the strongest inhibition of nitric oxide production (IC$\sub$50/ = 17 uM). And this inhibition was, at least partly, due to down-regulation of iNOS enzyme induction, not due to direct inhibition of iNOS enzyme activity.

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Effect of Scoparone on the Hepatic Microsomal UDPglucuronyltransferase Activity in Mice

  • Huh, Keun;Lee, Sang-Il;Park, Jong-Min
    • Archives of Pharmacal Research
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    • v.10 no.3
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    • pp.165-168
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    • 1987
  • The effect of scoparone on UDP glucuronyltransfearase in mouse hepatic microsomes was studied. After transment with scoparone, hepatic microsomal UDP glucuronyltransferase activity was increased with dose-dependent manner as compared to control. The V$_max$ value (control = 23.2 n moles/mg protein/min, scoparone = 31.2 n moles/ mg protein /min) without affecting the $K_m$ value (414 $\mu$M) for p-nitrophenol was increased by the scoparone treatment, and the pattern of kinetic studies for UDP-glucuronic acid was also similar to those of p-nitrophenol. Whereas, the hepatic microsomal UDP glucuronyl-transferase was not changed by the addition of scoparone in vitro. The results obtained suggest that the characteristics of increase in the enzyme activity may include induction of enzyme proteins.

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Induction of Phosphoenolpyruvate Carboxylase from Hydrilla verticillata Grown Under $CO_2$ Stress ($CO_2$ 스트레스에서 생장한 검정말로부터 Phosphoenolpyruvate carboxylase 유도)

  • Spencer, William E.;Kim, Yong-Jae
    • Korean Journal of Ecology and Environment
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    • v.38 no.4 s.114
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    • pp.454-460
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    • 2005
  • The goal of this research was to better understand the mechanism of environmental control of PEPC expression from Hydrilla verticillata grown under $CO_2$ stress. When PEPC concentration was plotted against $CO_2$ compensation point (${\Gamma}$), a linear increase in PEPC concentration was seen with a decrease in $CO_2$ compensation point. ${\Gamma}$ was measured on each day of the time course induction. Hydrilla plants exhibited an initially large decrease in ${\Gamma}$ between day 0 and day 1; thereafter, the ${\Gamma}$ declined in a fairly linear trend for the remaining 5 day time course. PEPC activity varied greatly over the time course. PEPC activity increased daily from day 0 through day 5 of the time course. Activity of PEPC increased almost 4-fold over the time course induction. The two upper bands with the approximately 100 kD mass are the PEPC subunits and PEPC was barely detectable in day 0 but increased through day 4 with day 5 being approximately equal to day 4. We provide evidence of increased PEPC protein concentration, and increased PEPC enzyme activity in as little as 24 h after beginning of induction.

Benzoyltransferase and Phenylacetyltransferase Activities in Cholestatic Rat Liver Induced by Common Bile Duct Ligation

  • Kim, Young-Jin;Kim, You-Hee
    • BMB Reports
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    • v.32 no.1
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    • pp.67-71
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    • 1999
  • We have investigated the effect of cholestasis on the closely related acyl-CoA:amino acid N-acyltransferase, benzoyltransferase, and phenylacetyltransferase activities in rat liver. Benzoyltransferase and phenylacetyltransferase activities in the liver cytosol, mitochondria, and microsome were investigated for a period of 42 d after common bile duct ligation. Both the mitochondrial and microsomal benzoyltransferases showed significant increase in their activities between the 1st and 7th day after common bile duct ligation, although the cytosolic benzoyltransferase activity did not show a significant change compared to the activities from the sham-operated control. The cytosolic phenylacetyltransferase activity showed a significant increase between the 1st and 2nd day, the mitochondrial activity showed a significant increase between the 2nd and 7th day, and microsomal activity showed a significant increase between the 1st and 7th day, respectively. Enzyme kinetic parameters of hepatic benzoyltransferase were analyzed using benzoyl coenzyme A as a substrate with the preparations from the 1st day post-ligation. Enzyme parameters of hepatic phenylacetyltransferase were also analyzed using phenylacetyl coenzyme A as a substrate with the preparations from the 2nd day post-ligation. The results indicated that although the $K_m$ values of these enzymes were about the same as the sham-operated control, the $V_{max}$ values of both enzymes increased significantly. These results, therefore, suggest that the biosynthesis of benzoyltransferase and phenylacetyltransferase has been induced in response to cholestasis.

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Tyrosine Hydroxylase Activity and mRNA in Rat Locus Coeruleus and Adrenals Following Chronic Ethanol Treatment and Acute Cold Stress

  • Lee, Yong-Kyu;Park, Dong-Ha
    • BMB Reports
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    • v.29 no.5
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    • pp.393-397
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    • 1996
  • Sprague-Dawley male rats (150 g) were chronically treated with 5 v/v % ethanol admixed with nutritionally complete liquid diet and fed ad libitum for 3 weeks. Controls were pair fed with the isocaloric sucrose liquid diet. One half of each group was exposed to cold stress at $4^{\circ}C$ either for 24 h (for determination of mRNA by in situ hybridization) or for 48 h (for determination of enzyme activity). Chronic ethanol treatment (ethanol) did not affect tyrosine hydroxylase (TH) mRNA level in locus coeruleus (LC) of brain and adrenal medulla (AM) compared to controls. Cold stress showed strong increase of TH mRNA level in LC and AM compared to controls. Pretreated ethanol reduced the increased TH mRNA level by cold stress in LC and AM. Ethanol did not affect TH activity in LC and adrenal glands (adrenals). Cold stress increased TH activity in LC but not in adrenals. Pretreated ethanol did not reduce the increased TH activity by cold stress in LC but this result was not shown in adrenals. It is suggested that ethanol does not affect the message level and enzyme protein level for TH in LC and AM in normal rat. It is also hypothesized that pretreated ethanol reduces the magnitude of acute cold stress response, that is induction of TH mRNA in LC and AM, and does not reduce the increased TH enzyme protein that is also acute cold stress response in LC.

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Effects of Oxidative Stress on Apoptosis and Antioxidant Enzyme Levels

  • Kim, Choonmi;Lee, Ji-Young
    • Biomolecules & Therapeutics
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    • v.4 no.4
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    • pp.402-407
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    • 1996
  • Effects of oxidative stress on the induction of apoptosis and the activity of antioxidant enzymes were investigated in HL-60 cells using $H_2O$$_2$and cisplatin which generate oxygen species in the cell. Various concentrations of oxidants were treated to cells and at different incubation time, cells were harvested for assays. Cell viability, morphology by propidium iodide staining and DNA fragmentation by agarose gel electrophoresis were observed to determine whether they induce apoptosis. The activity of antioxidant enzymes such as superoxide dismutase and catalase was also measured to evaluate the cellular response to the oxidative damage. The results are as follows: $H_2O$$_2$ induced apoptosis at 10 $\mu$M after 6h incubation, while it took 12h for cisplatin. Both oxidants induced the superoxide dismutase activity at a tolerable low concentration. However, at a concentration which causes apoptotic cell death, the enzyme level was dropped markedly at first and then recovered to the normal level after which it declined again, probably due to cell death. On the other hand, changes in the activity of catalase were not significant at most concentrations except the statistically significant decrease at 24h after 10 $\mu$M-$H_2O$$_2$treatment. In this study, $H_2O$$_2$- and cisplatintreated cells showed similar results in apoptotic response and enzyme activities, suggesting that anticancer activity of cisplatin may be related, at least in part, to the production of oxygen free radicals.

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The Effect of Aspalatone, a New Antithrombotic Agent, on the Specific Activity of Antioxidant Enzyme in the Rat Blood

  • Kim, Chin;Koo, Chang-Hui;Choi, Dong-Young;Cho, Yong-Joon;Choi, Jae-Ho;Im, Doo-Hyeon;Jhoo, Wang-Kee;Kim, Hyoung-Chun
    • Archives of Pharmacal Research
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    • v.19 no.5
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    • pp.348-352
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    • 1996
  • The antioxidant efficacy of aspalatone, a new antithrombotic agent, has been recognized in the neurotoxic model and in the cardiotoxic model in proliminary studies. We examined the specific activity of antiosidnat enzyme in the rat blood following administrations of aspirin, maltol, aspirin together with maltol, salicylmaltol (major metabolite of aspalatone) and aspalatone, respectively. Our assessment showed that salicylmaltol, maltol, aspalatone enhanced antiperoxidative activity. In addition, neither aspirin nor combination of aspirin and maltol, showed any significant effect on the activity of antioxidant enzyme. Because $H_{2}$$O_{2}$ accumulation may stimulate the thrombogenesis in blood, the result suggests that the induction of blood antiperoxidative activity produced by aspalatone may have beneficial effects on the thrombogenesis.

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