• Journal of Microbiology and Biotechnology
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• 제1권4호
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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.

• Archives of Pharmacal Research
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• 제13권1호
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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.

• 한국연초학회지
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• 제20권1호
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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$^{\circ}C$).

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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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.

• Archives of Pharmacal Research
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• 제10권3호
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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.

• 생태와환경
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• 제38권4호통권114호
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• pp.454-460
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• 2005

본 연구는 $CO_2$ 스트레스 하에서 생장한 검정말로부터 PEPC 단백질 발현에 대한 환경조절 기작을 잘 이해하도록 하는데 그 목적이 있다. PEPC 농도와 $CO_2$ 보상점과의 관계에서, PEPC농도는 $CO_2$ 보상점 (${\Gamma}$)이 감소할 때 직선상으로 증가하였다. ${\Gamma}$를 유도하는 과정 동안 매일 측정하였다. 검정말은 초기 0일과 1일 사이에 ${\Gamma}$를 급감시키며, 이후 5일까지는 완만한 감소를 나타내었다. PEPC 활성은 0일에서 5일까지 유도 과정 동안 매일 증가하였으며, 5일 동안 4배 이상 증가하였다. PEPC는 100 KD으로 상부에 두 개의 band를 나타내었다. 본 연구의 결과 유도 시작 후 24시간 내에 PEPC 단백질과 효소활성이 증가하였다.

• BMB Reports
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• 제32권1호
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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.

• BMB Reports
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• 제29권5호
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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.

• Biomolecules & Therapeutics
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• 제4권4호
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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.

• Archives of Pharmacal Research
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• 제19권5호
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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.