• Journal of Korean Society of Environmental Engineers
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• v.22 no.3
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• pp.417-427
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• 2000

The substrate metabolism and bacterial population in an anaerobic digestion with the submerged separation membrane were investigated by using a laboratory-scale reactor at the hydraulic retention time(HRT) 1.0 and 0.5 day. The removal efficiencies of carbohydrate at the HRT 1.0 and 0.5 day were 99.8~99.9% and 98.0~99.6%, respectively. After the 58 days, the mixed liquor volatile suspended solids(MLVSS) concentration at the HRT 1.0 and 0.5 day were approximately 6,050 and 7,750 mg/L, respectively. According to the measurement by the most probable number(MPN) method, the numbers of acidogenic bacteria, $H_2$-utilizing and acetate-utilizing methc.nogenic bacteria were found to be $10^9$, $10^7{\sim}10^8$ and $10^6{\sim}10^8MPN/mL$, respectively. The composition of $CH_4$ in the produced gas was 46~50%. It is suggested that sulfate-reducing bacteria $10^7{\sim}10^8MPN/mL$ play an important role in producing $H_2$ and acetate in sulfate-depleted environment.

• Applied Biological Chemistry
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• v.37 no.3
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• pp.203-209
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• 1994

When $^{14}C-{\alpha}-endosulfan$ was incubated with carp liver, kidney and gut preparations, it was metabolized to water soluble and organosoluble metabolites. In an in vitro test, endosulfan was converted to endosulfan ${\alpha}-hydroxyether$ (EHE), endosulfan alcohol (EA) and endosulfan ether (EE). The addition of NADPH resulted in rapid conversion of endosulfan to the metabolites in 105,000 g soluble fraction and microsomes. However, the rate of metabolism of endosulfan in liver, kidney and gut supplemented with NADPH as a cofactor was higher in the 105,000 g soluble fraction than that in the microsomes of carp under incubation conditions. The enzymes probably involved in the metabolism of endosulfan include the glutathione S-transferase (GST) and the mixed function oxidases (MFO), based on the evidence that addition of either GSH or NADPH increased the degradation of endosulfan.

• BMB Reports
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• v.28 no.2
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• pp.162-169
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• 1995

Rat brain succinic semialdehyde dehydrogenase (EC 1.2.1.24 SSADH) activity was detected in mitochondrial, cytosolic and microsomal fractions. Brain mitochondrial soluble SSADH was purified by ammonium sulfate precipitation, DEAE Sephacel, and 5'-AMP Sepharose 4B affinity chromatography. The purified enzyme was shown to consist of four identical subunits, and the molecular weight of a subunit was 55 kD. The $K_m$ for short chain aliphatic aldehydes and aromatic aldehydes were at the $10^{-3}M$ level but that for succinic semialdehyde was 2.2 ${\mu}M$. Either $NAD^+$ or $NADP^+$ can be used as a cofactor but the affinity for $NAD^+$ was 10 times higher than that for $NADP^+$. The brain cytosolic SSADH was also purified by ammonium sulfate precipitation, DEAE Sephacel, Blue Sepharose CL-6B and 5'-AMP Sepharose 4B affinity chromatography and its Km for short chain aliphatic aldehydes was at the $10^{-3}$ level but that for succinic semialdehyde was 3.3 ${\mu}M$. $NAD^+$ can be used as a cofactor for this enzyme. We suppose that both enzyme might participate in the oxidation of succinic semialdehyde, which is produced during GABA metabolism. The activity of both cytosolic and mitochondrial SSADH was markedly inhibited when the concentration of succinic semialdehyde was high. The reciprocal plot pattern of product inhibition and initial velocity indicated a sequential ordered mechanism for mitochondrial matrix SSADH. Chemical modification data suggested that amino acid residues such as cysteine, serine and lysine might participate in the SSADH reaction.

• Korean Journal of Crystallography
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• v.4 no.2
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• pp.100-104
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• 1993

Reduced glutathione (GSH) plays a vital role in the metabolism of all cells. Glutathions, a tripeptide cowfosed of glutamic acid, cysteane, and gtycina is synthesized by two synthesized reutions. The first is catalyzed by Y-glutamylcysteine synthetase (GSH-I) and the second by glutathione synthetase (GSH-ll). The glutathione biosynthetic pathway of E. coziis mainly controlled by nonallosteric feedback inhibition of GHS-I by GSH. Determination of the three-dimensional structure of GSH-I by X-ray crystallography is necessary in order to understand the structure-function relationship at the molecular level. As the (irst step toward its structure determination, crystallization of 5. coli V-glutamylcystfine synthetase (GSH-I) has been achived using the hanging drop vapor diffusion method and capillaw method. Crystals of GSH-I have been grown from ammonium sulfate solution. The crystals grew at room temperature within 10 days to dimensions of 0.2 m x 0.2 m x 0.2 ml by hanging drop vapor diffusion method and diffracted to about 4 A resolution using synchrotron X-rays. Another crystal, grown by the capillary method to dimensions of 0.25 mm x 0.25 mm x 0.3 mm within 40 days, diffracted to about 4 A resolution using X-rays from a rotating anode.

• Korean Journal of Food Science and Technology
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• v.37 no.1
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• pp.90-94
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• 2005

Absorption, metabolism, and tissue concentrations of quercetin were examined and compared in mice and rats after oral administration of quercetin at 50 or 100 mg/kg. Quercetin was absorbed quickly in mice and reached maximum plasma concentration in I hr post-administration, and declined sharply after 4 hr. Plasma concentration of isorhamnetin, a major metabolite, also increased sharply, indicating rapid metabolic conversion, but elevated level was maintained longer than that of quercetin. Quercetin and isorhamnetin were found predominantly in glucuronide/sulfate-conjugate forms in both mice and rats. Tissue concentrations of quercetin and isorhamnetin in mice and rats were in the order of liver>kidney>spleen>plasma both 1 and 6 hr postadministration. These results show that quercetin is absorbed in mice after oral feeding and quickly metabolized into isorhamnetin as demonstrated in humans and other animal species. The results also can be used to explain various pharmacological activities reported in mouse models.

• Journal of The Korean Society of Grassland and Forage Science
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• v.42 no.3
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• pp.195-200
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• 2022

Sulfur is an essential element in plants, including amino acids, vitamin synthesis, and acting as an antioxidant. However, the interaction between endogenous sulfur and proline synthesis has not been yet fully documented. White clover (Trifolium repens L.) is known as a species highly sensitive to sulfate supply. Therefore, this study aimed to elucidate the role of sulfur in regulating proline metabolism in relation to ammonia detoxification and hydrogen peroxide (H₂O₂) accumulation in white clover. The detached leaves of white clover were immersed in solution containing different concentration of sulfate (0, 10, 100, and 1000 mM MgSO₄). As MgSO₄ concentrations were increased, the concentration of H₂O₂ increased up to 2.5-fold compared to control, accompanied with H₂O₂ detection in leaves. Amino acid concentrations significantly increased only at higher levels (100 and 1000 mM MgSO₄). No significant difference was observed in protein concentration. Proline and ∆^1-pyrroline-5-carboxylate (P5C) concentrations slightly decreased at 10 and 100 mM MgSO₄ treatments, whereas it rapidly increased over 1.9-fold at 1000 mM MgSO₄ treatment. Ammonia concentrations gradually increased up to 8.6-fold. These results indicate that exogenous sulfur levels are closely related to H₂O₂ and ammonia synthesis but affect proline biosynthesis only at a higher level.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1998.11a
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• pp.191-191
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• 1998

It has been hypothesized that formation of oxygen-derived free radicals may play an important part in ischemically induced tissue injury. In this study, the effects of vitamin C treatment on hepatic reperfusion model were investigated. Livers isolated from 18 hrs fasted rats were subjected to low flow hypoxia (1 $m\ell$/g liver/min, for 45min) followed by reoxygenation (for 30min). The perfusion medium used was Krebs-Henseleit bicarbonate buffer (KHBB, pH 7.4) and vitamin C (0.25, 0.5, 1.0 and 2.0 mM) was added to perfusate. 7-Ethoxycoumarin was used as substrate of phase and metbolism. After hypoxia oxygen consumption significantly dropped but vitamin C 0.25, 0.5 and 1.0 mM treatments restored oxygen consumption to the level of control group. LDH and lipid peroxidation were not changed in all experimental groups. Oxidation, phase metabolism, significantly decreased following hypoxia but improved during reoxygenation. Vitamin C 0.25 mM treatment significantly improved the oxidation of 7-ethoxycoumarin during hypoxia and reoxygenation, but the oxidation significantly decreased by vitamin C 2.0 mM treatment. Similarly, sulfate conjugation decreased in hypoxic group, but this decrease was inhibited by vitamin C 0.25, 0.5 and 1.0 mM treatments. Our findings suggest that hypoxia/reoxygenation diminishes hepatic drug metabolizing function, vitamin C at concentration of 0.25-1.0 mM ameliorates but at higher concentration aggravates these hypoxia/reoxygenation-induced changes.

• Journal of the Korean Chemical Society
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• v.54 no.5
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• pp.567-572
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• 2010

5,10-Methenyltetrahydrofolate synthetase from chicken liver was purified through 30-70% ammonium sulfate fractionation, Q Sepharose Fast Flow anion exchange and Source 15Phe hydrophobic interaction chromatography. Specific activities of cell extract, ammonium sulfate, Q Sepharose Fast Flow and Source 15Phe were 0.0085, 0.031, 0.80 and 1.27 U/mg, respectively. Purification fold activities of cell extract, ammonium sulfate, Q Sepharose Fast Flow and Source 15Phe were 1, 3.7, 94.1 and 149.4, respectively. HPLC gel permeation chromatography and SDS-polyacrylamide electrophoresis experiments indicated that the enzyme is a monomeric protein with a molecular weight of 22.8 kDa. Km for 5-methyl THF and Mg-ATP were $7.1\;{\mu}M$ and $63\;{\mu}M$, respectively. Optimum temperature and pH were $30^{\circ}C$ and 6.0, respectively. The data for metal ion specificity and stoichiometry showed that the maximum activity was obtained with a 1:l. ratio of $Mg^{2+}$. The ATP and Km values increased in the order of MgATP, MgCTP, MgUTP and MgGTP, and the maximum activities also decreased in the same order, indicating MgATP as the most efficient substrate. The enzyme was chemically modified only by tetranitrometane and 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide, indicating that tyrosine and carboxylate are present in the active site.

• Archives of Pharmacal Research
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• v.23 no.6
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• pp.620-625
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• 2000

The mechanisms of liver injury from cold storage and reperfusion are not completely under-stood. The aim of the present study was to investigate whether the inactivation of Kupffer cells (KCs) by gadolinium chloride ($GdCl_3$) modulates ischemia-reperfusion injury in the rat liver. Hepatic function was assessed using an isolated perfused rat liver model. In livers subjected to cold storage at $4^{\circ}C$ in University of Wisconsin solution for 24 hrs and to 20 min rewarm-ing ischemia, oxygen uptake was markedly decreased, Kupffer cell phagocytosis was stimulated, releases of purine nucleoside phosphorylase and lactate dehydrogenase were increased as compared with control livers. Pretreatment of rats with $GdCl_3$) , a selective KC toxicant, suppressed kupffer cell activity, and reduced the grade of hepatic injury induced by ischemia-reperfusion. While the initial mixed function oxidation of 7-ethoxycoumarin was not different from that found in the control livers, the subsequent conjugation of its meta-bolite to sulfate and glucuronide esters was suppressed by ischemia-reperfusion, CdCl$_3$restored sulfation and glucuronidation capacities to the level of the control liver. Our findings suggest that Kupffer cells could play an important role in cold/warm ischemia-reperfusion hepatic injury.

• Archives of Pharmacal Research
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• v.20 no.5
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• pp.459-464
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• 1997

The human liver cDNA clone UDPGTh2, encoding a liver UDP-glucuronosyltransferase (UDPGT) was isolated from a .gamma. gt 11 cDNA library by hybridization to mouse transferase cDNA clone, UDPGTm1. UDPGTh2 encoded a 529 amino acid protein with an amino terminus membrane-insertion signal peptide and a carboxyl terminus membrane-spanning region. There were three potential asparagine-linked glycosylation sites at residues 67, 68, and 315. In order to obtain UDPGTh2 protein encoded from cloned human liver UDP-glucuronosyltransferase cDNA, the clone was inserted into the pSVL vector (pUDPGTh2) and expressed in COS 1 cells. The presence of a transferase with Mr-52,000 in transfected cells cultured in the presence of $[^{35}S]$ methionine was shown by immunocomplexed products with goat antimouse transferase IgG and protein A-Sepharose and analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. The expressed UDPGT was a glycoprotein as indicated by electrophoretic mobility shift in Mr-3,000-4,000 when expressed in the presence of tunicamycin. The extent of glycosylation was difficult to assess, although one could assume that glycosyl structures incorporated at the level of endoplasmic reticulum were always the core oligosaccharides. Thus, it is likely that at least two moieties inserted can account for the shift of Mr-3,000-4,000. This study demonstrates the cDNA and deduced amino acid sequence of human liver UDP-glucuronosyltransferase cDNA, UDPGTh2.