• Title/Summary/Keyword: L-citrulline

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High Production of L-Crnithine by L-Citrulline Auxotroph of Brevibacterium ketoglutamicum : PART I : Selection of Stable Mutant Strains with Less Revertant Formation (Brevibacterium ketoglutamicum을 이용한 L-Ornithine 생산 연구 PART I : L-Ornithine 생산 고역가 안정화 균주 선발)

  • 류욱상;장형욱;이홍원;정준기;장순재;유연우;박영훈
    • Microbiology and Biotechnology Letters
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    • v.27 no.4
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    • pp.292-297
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    • 1999
  • Overproduction of L-ornithine by mutant strains isolated from Brevibacterium detoglutamicum BK1046 was investigated. The strain was a L-ornithine auxotroph and exhibited culture instability during fermentation. Through a sequential screening effort, a highly stable strain with lless revertant formation was finally selected and designated B. ketoglutamicum BK52 (KCTC0141BP). It prouduced L-ornithine at a high concentration (above 9 g/L) independent of subculture or cultivation time, and also had a very low tendency of revertant formation. In a long-term storage, this strain maintained its cell stability and productivity of L-ornithine to a reasonable range.

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Purification and Some Properties of Arginine Deiminase in Euglena gracilis Z (Euglena gracilis Z로부터 Arginine Deiminase의 정제 및 그의 특성)

  • Park, Bong-Sun;Hirotani, Aiko;Nakano, Yoshihisa;Kitaoka, Shozaburo
    • Journal of the Korea Organic Resources Recycling Association
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    • v.1 no.1
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    • pp.115-125
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    • 1993
  • In Euglena gracilis arginine deiminase was located in the mitochondrial matrix. The highly purified enzyme required $Co^{2+}$ for the enzyme reaction with the $K_m$ value of 0.23 nM, and its optimum pH was 9.7 to 10.3. The molecular weight of the native enzyme protein was 87,000 by gel filtration, and SDS-acrylamide gel electrophoresis showed that the enzyme consisted of two identical subunits with a molecular weight of 48,000. Euglena arginine deiminase was inhibited by sulfhydryl inhibitors, indicating that a sulfhydryl group is involved in the active center of the enzyme. It exhibited negative cooperativity in binding with arginine. $L-{\alpha}-amino-{\beta}-guanidino-propionate$, D-arginine, and L-homoarginine strongly inhibited the enzyme while ${\beta}-guanidinopro-pionate$, ${\gamma}-guanidinobutyrate$, and guanidinosuccinate did not. Considerable inhibition was also observed with citrulline and ornithine. We discuss the effects of the unique properties of the Euglena arginine deiminase on the regulation of arginine metabolism in this protozoon.

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Effect of Ginsenoside on Basal and Nitro-L-Arginine Suppressed Nitric Oxide Production in Rat Kidney

  • Kim, Hye-Young;Han, Sang-Won
    • Biomolecules & Therapeutics
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    • v.2 no.2
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    • pp.131-135
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    • 1994
  • The effect of ginsenoside (GS) from Panax ginseng on basal and nitro-L-arginine suppressed nitric oxide (NO) production was studied in rat kidney. NO production was determined by conversion to [$^{14C}$]=L-citrulline from [$^{14C}$]-L-arginine both in whole kidney and three renal segments; glomerulus, cortex excluding glomerulus (cortex-) and medulla. Nitro-L-arginine (total dose of 30 mg/kg/3 days, i.p.) significantly reduced NO production in whole kidney, which was prevented by GS pretreatment (30 mg/kg/3 days, i.p.). Relative high dose of GS (120 mg/kg/4 days, i.p..) selectively increased NO production in glomerulus and cortex-. Protein content, on wet weight basis, in cortex- and glomerular DNA content were significantly reduced by GS. Our results confirm the existence of constitutive nitric oxide synthase in kidney and it seems that target nephron segment for volume expansion due to GS'NO-mediated vasodilation and for NO production stimulated by GS is cortex including glomerulus.lus.

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Screening Test(I) of Several Antipsychotic Agents on NO Formation (수종 정신병치료제들의 NO형성에 대한 검색(I))

  • Lee, Jong-Hwa;EI-fakahany, Esam E.
    • The Korean Journal of Pharmacology
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    • v.30 no.3
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    • pp.343-349
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    • 1994
  • A number of neurological syndromes(e.g. tardive dyskinesia) are developed as a consequence of chronic treatment with neuroleptics or antipsychotic agents. Despite the long and succesful use of phenothiazine derivatives and related agents in the treatment of certain states of mental disease, the mechanisms of these drugs are still poorly understood. One current hypothesis from extensive reviews is that these compounds might significantly interfere with the cyclic nucleotide system in brain (Levin and Weiss, 1977; Nowicki et al., 1991; Haley et al., 1992). Nitric oxide (NO), one of an interesting messenger molecule and aberrant transmitter, is believed to play a important role in biological functions of cyclic nucleotides in nervous system. It has been reported that calcium-dependent NO synthesis in endothelial cytosol is mediated by calmodulin which is supposed to be tightly related to pharmacological actions of antipsychotic agents. In the present study, the effect of several antipsychotic agents on the activity of NO synthesis and formation of cyclic GMP were investigated. These agents inhibited both the formation of $[^3H]L-citrulline$ and that of $[^3H]cyclic$ GMP by concentration-dependent manner, and their inhibiting patterns are so similar to that of calmodulin antagonist.

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Role of Nitric Oxide in Pepsinogen Secretion from Rat Gastric Chief Cells

  • Sung, Dae-Suk;Seo, Dong-Wan;Choi, Don-Woong;Ahn, Seong-Hoon;Hong, Sung-Youl;Lee, Hoi-Young;Han, Jeung-Whan;Lee, Hyang-Woo
    • Biomolecules & Therapeutics
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    • v.7 no.2
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    • pp.105-111
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    • 1999
  • Nitric oxide (NO), a cellular messenger synthesized from L-arginine by NO synthase (NOS, EC.1.14.13.39), is considered to be a regulator of gastric secretion. In the present study, the role of NO in the regulation of exocrine secretion was investigated in rat gastric chief cells. Treatment of chief cells with carba-chol resulted in an increase in the arginine conversion to citrulline, the amount of $NO_{x}$, the release of pepsine-gen, and the level of cGMP Especially, carbachol-stimulated increase of arginine to citrulline transformation, the amount of $NO_{x}$, cGMP level and the release of pepsinogen were partially reduced by the natural NOS inhibitor, $N^{G}$-monomethyl-L-arginine (MMA) and $N^{G}$, $N^{G}$-dimethyl-L-arginine (DMA). Furthermore, MMA- and DMA-induced decrease of pepsinogen secretion showed dose-dependent patters. Activation of NOS is one of the early events in receptor-mediated cascade of reactions in gastric chief cells and NO, not completely, but partially mediates gastric secretion. Agonist-stimulated pepsinogen secretion in chief cells has been considered to be mediated in adenosine 3',5'-cyclic monophosphate pathway and/or guanosine 3', 5'-cyclic monophosphate (cGMP) pathway. Taken together, the above results suggest that partial decrease of exocrine secretion following treatment of NOS inhibitor may result from the inactivation of NOS and subsequent guano- late cyclase, and NO/cGMP pathway may play a pivotal role in exocrine secretion.

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High Production of L-Ornithine by L-Citrulline Auxotroph of Breviabcterium ketoglutamicum : PART II : Production of L-Ornithine by Controlled Feeding of L-Arginine (Brevibacterium ketoglutamicum을 이용한 L-Ornithine 생산 연구 PART II : L-Arginine 제한공급에 의한 :-Ornithine 유가식 발효생산)

  • 류욱상;장형욱;이홍원;정준기;장순재;유연우;박영훈
    • Microbiology and Biotechnology Letters
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    • v.27 no.4
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    • pp.327-332
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    • 1999
  • A highly productive fed-batch fermentation process was developed for the production of L-ornithine by using a new stabilized strain, Breviabcterium ketoglutamicum BK52. Fed-batch cultures with a continuous feeding of the complex medium were conducted on various operating conditions. The optimal concentration of phosphate in the complex medium was 2.1g/L. The optimal feeding rate of L-arginine was 0.028g/L/hr. The optimal feeding point of the complex medium was determined to be at 40 OD of the cell mass. The final L-ornithine concentrations within 64hrs of cultivation in 5 and 50 liter fermenters were 73g/L and 71g/L, respectively. The maximum overall L-ornithine productivity was 1.14g/L/hr which was about 2 times higher than that of the conventional fed-batch culture with intermittent feeding. The overall productivity of the fermentation system is remarkably improved by employing the optimized conditions, and it offers a significant potential for industrial application.

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Acid Secretion and Nitric Oxide Synthase Activity in Gastric Glands Following Hypoxia/Reoxygenation and Acidosis (Hypoxia/Reoxygenation과 Acidosis가 위선세포에서 위산분비와 NO Synthase 활성에 미치는 영향)

  • Kim Hye-Young;Kim Kyung-Hwan
    • The Korean Journal of Pharmacology
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    • v.31 no.1 s.57
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    • pp.75-84
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    • 1995
  • Acid secretion and NO synthase activity were determined in isolated gastric glands following hypoxia/reoxygenation and acidosis to investigate the involvement of NO in acid secretion. Isolated gastric glands were exposed to hypoxia (30 min)/reoxygenation (1 h) and/or to acidosis (pH 6.0 and 4.0). Acid secretion was measured by the ratio of $[^{14}C]-aminopyrine$ accumulation between intra- and extraglands. NO synthase activity was determined by percent conversion to $[^{14}C]-citrulline\;from\;[^{14}C]L-arginine$, a precursor of NO. The results indicate that dibutyryl cAMP stimulated acid secretion dose-dependently but had no effect on NO synthase activity in basal gastric glands. Hypoxia/reoxygenation significantly suppressed acid secretion both in unstimulated and stimulated gastric glands, which was exaggerated by acidosis. Constitutive NO synthase, activity, not responded to dibutyryl cAMP, was also inhibited by hypoxia/reoxygenation and acidosis. In conclusion, pathologic state of gastric mucosa such as hypoxia/reoxygenation and acidosis suppresses both acid secretion and NO release but the role of NO in acid secretion stimulated by dibutyryl cAMP in basal gastric glands is not significant.

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Identification and Characterization of Nitric Oxide Synthase in Salmonella typhimurium

  • Choi, Don-Woong;Oh, Hye-Young;Hong, Sung-Youl;Han, Jeung-Whan;Lee, Hyang-Woo
    • Archives of Pharmacal Research
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    • v.23 no.4
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    • pp.407-412
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    • 2000
  • The presence of the nitric oxide synthase (NOS) enzyme from Salmonella typhimurium (S. typhimurium) was identified by measuring radiolabeled L-$[^3H]$citrulline and NO, and Western blot analysis. NOS was partially purified by both Mono Q ion exchange and Superose 12HR size exclusion column chromatography, sequentially. The molecular weight of NOS was estimated to be 93.3 kDa by Western blot analysis. The enzyme showed a significant dependency on the typical NOS cofactors; an apparent Km for L-arginine of 34.7 mM and maximum activity between $37^{\circ}C$ and $43^{\circ}C$. The activity was inhibited by NOS inhibitors such as aminoguanidine and $N^{G}$ $N^{G}$-dimethyl-L-arginine. taken together, partially purified NOS in S. typhimurium is assumed to be a different isoform of mammalian NOSs.OSs.

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Amino Acid, Amino Acid Metabolite, and GABA Content of Three Domestic Tomato Varieties

  • Ahn, Jun-Bae
    • Culinary science and hospitality research
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    • v.22 no.6
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    • pp.71-77
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    • 2016
  • To determine the nutritional value of domestic tomatoes, the levels of amino acids, amino acid metabolites, and the bioactive compound ${\gamma}-aminobutyric-acid$ (GABA) were analyzed in three domestic tomato varieties (Rafito, Momotaro, and Medison). Eighteen free amino acids were found, and total free amino acid content was 3,810.21~4,594.56 mg/100 g (dry weight). L-glutamic acid (L-Glu) was the most abundant amino acid, ranging from 1,866.60 mg/100 g for Momotaro to 2,417.45 mg/100 g for Medison. The next most abundant amino acids were L-glutamine (L-Gln) and L-aspartic acid (L-Asp). The three tomato varieties had a good balance of all the essential amino acids except tryptophan. Total essential amino acid content was 274.26~472.71 mg/100 g (dry weight). The following amino acid metabolites were found: L-carnitine (L-Car), hydroxylysine (Hyl), o-phosphoethanolamine (o-Pea), phosphoserine (p-Ser), ${\beta}-alanine$ (${\beta}-Ala$), N-methyl-histidine (Me-His), ethanolamine (EtNH2),and L-citrulline(L-Cit). Large quantities of GABA were found in all three varieties: 666.95-868.48 mg/100g (dry weight). These results support the use of these tomato varieties as nutritious food materials.

Stimulatory Effect of Ginseng Saponin on Endogenous Production of Nitic Oxide

  • Kim, Hye-Young
    • Proceedings of the Ginseng society Conference
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    • 1998.06a
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    • pp.199-207
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    • 1998
  • Ginseng saponin (G5) purified from Panax ginseng, increase renal blood flow in rats. Nitric oxide (NO) is thought to be a substance endogenously released by G5 in preconstricted lungs and cultured endothelial cells. The present study aims to determine whether G5 could stimulate endogenous 1'elease of NO in rat kidney and urine levels of the stable NO metabolites, nitrite (NO,) and nitrate (NO,) and urinary COMP levels were measured 8 hr after a single intraperitoneal injection of GS (200 mg/kg) Into rats. The effects of the WO synthesis inhibitor, Nu-nitro-L-arginine methyl ester, .1nd the NO precursor, L-arginine, on the G5-induced changes were also determined. The activity of NO synthase, as determined by conversion of ('"C)-L-arginine to ('"C)-L-citrulline, in whole kidney, glomeruli and cortical tubules were also investigated. A single injection of GS resulted in endogenous production of NO as reflected by increase in serum and urine levels of N021N03 and urinary cGMP levels, which were inhibited by the addition o ( N-nitro-L-arginine methyl ester and restored fly L-arginine. GS also stimulated the activity of NO synthase in whole kidney as well as glomeruli and cortical tubules, and Nu-nitro-L-arginine methyl tilter significantly prevented this increase. In conclusion, GS stimulates endogenous NO production and thus, may play a protective role 1 11 the kidney by modulating renal blood flow.

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