• Title/Summary/Keyword: L-glutamate

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Production of L-Ornithine by Citrulline Auxotrophic Mutants of Glutamate-Producing Bacteria

  • CHOI, DAE KEON;WUK SANG RYU;BONG HYUN CHUNG;SOO WAN NAM;YOUNG HOON PARK
    • Journal of Microbiology and Biotechnology
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    • v.2 no.2
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    • pp.102-107
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    • 1992
  • For the purpose of producing L-ornithine by microbial fermentation, mutant strains were developed from glutamate-producing bacteria by mutagenesis using N-methyl-N'-nitro-N-nitrosoguanidine (NTG) and UV irradiation. Brevibacterium ketoglutamicum BK1046, a L-citrulline auxotroph which is also resistant to arginine hydroxamate (Arghx), was isolated and selected as the best producer of L-ornithine. This strain was capable of producing L-ornithine at a concentration of 24 g/l after 69 hours of cultivation in the 21 jar fermentor. The optimum supplementary level of L-arginine, a substitute for L-citrulline, was found to be about 0.2 g/l in the shake-flask fermentation.

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Enhanced Production of L-Aspartate ${\beta}-Decarboxylase$ by Nitrogen Source in Pseudomonas dacunhae

  • Kim, Dong-Chung;Lee, Sung-Dong;In, Man-Jin
    • Journal of Applied Biological Chemistry
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    • v.49 no.3
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    • pp.106-109
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    • 2006
  • Improvement of L-asparate ${\beta}-decarboxylase$ production from Pseudomonas dacunhae ATCC 21192 was attempted by optimizing fermentation conditions. Optimum carbon and nitrogen sources for cell growth and enzyme production were determined. L-Glutamate (2%) was the most suitable carbon source, and D-glucose, D-glycerol and fumarate repressed enzyme production. Yeast extract (2%) was the most effective as nitrogen source. A slight change of pH to 6.5 from medium pH resulted in a meaningful increase in the production of enzyme. The production of the enzyme was highly improved by using 2% yeast extract and 2% L-glutamate in culture media. Maximum L-asparate ${\beta}-decarboxylase$ activity reached up to over 24 U/mL-broth by 15 h flask fermentation.

Effects of Gultamate Synthesized during Photorespiration on Photosynthetic Carbon Metabolism (광호흡 과정에서 생성된 Glutamate가 광합성 탄소대사에 미치는 영향)

  • 이인철
    • Journal of Plant Biology
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    • v.31 no.4
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    • pp.277-288
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    • 1988
  • The effects of ammonium ion and glutamate on CO2 fixation abilities and related carbon metabolism were investigated in pea (Pisum sativum L. cv. Sparkle) leaf discs under conditions favoring photorespiration (21% O2, 0.03% CO2) and nonphotorespiration (5% O2, 0.03% CO2). A concentration of more than 10 mM of NH4+ decreased the photosynthetic CO2 fixation and those inhibitory effects were more remarkable in 21% O2 than in 5% O2 conditions. The effect of glutamate on CO2 fixation was found to be independent of the O2 level, as glutamate increased the CO2 fixation under both 21% and 5% O2 conditions. L-methionine-dl-sulfoximine, an irreversible inhibitor of glutamate synthetase, however, inhibited the CO2 fixation markedly under 21% O2, but did not affect it under 5% O2 conditions. The treatment with NH4+ elevated the relative amounts of 14C incorporated into soluble components from 14CO2 with no relation to O2 levels, while glutamate increased 14C into insoluble components and neutral sugars. Glutamate, especially, seemed to stmulate the biosynthesis of starch under 5% O2 condition. These results indicated that NH4+ stimulated the degradation of sugar or starch and this proposal was confirmed by the increasing of pyruvate kinase activity in leaf discs treated with ammonium ion.

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Genetic Analysis of Recombinants by Interspecific Protoplast Fusion of Coryneform Bacteria and Their L-glutamate & L-glutamine Production (Corynebacterium 세균의 이종간 원형질체 융합에 의한 재조합주의 유전학적 분석과 L-glutamate와 L-glutamine 생성)

  • 백선영;이혜경;최순영;김종욱;이세배;임번삼;민경희
    • Microbiology and Biotechnology Letters
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    • v.18 no.3
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    • pp.296-300
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    • 1990
  • For interspecific portoplast fusion, Brevibacterium flauum lOAHR (Rifr axg his) and Corynebacterium glutamicum 11TS ($Sm-r$ trp) were induced by UV and NTG treatment. The protoplast fusion frequency between E. flavum XOAHR and C. glutamicum llTS was $3.7\times 10^{-6}$ with the lysozyme treatment (300 P $\mu g$ml) for 18 hrs. Genotypes of recombinants were analized as FMM ($Rif^r\; Sm^r$), FA (Rift $Sm^r$ arg), FH ($Rif^r\; Sm^r$ his), FT ($Rif^r\; Sm^r$ trp), FAH ($Rif^r\; Sm^r$ arg trp), FAT ($Rif^r\; Sm^r$ arg trp), and FAHT ($Rif^r\; Sm^r$ arg his trp). FAH 1 produced 12 fold of glutamate production compared to parental type, E. flauum 10AHR. In glutamine productivity, it produced 2.6 fold to parental type, C. glutamicum 11TS. Production of glutamate or glutamine by recombinants was involved in the specific activities of glutamate dehydrogenase (GDH) and glutamine synthetase (GS), respectively.

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Effect of Various Pathological Conditions on Nitric Oxide Level and L-Citrulline Uptake in Motor Neuron-Like (NSC-34) Cell Lines

  • Shashi Gautam;Sana Latif;Young-Sook Kang
    • Biomolecules & Therapeutics
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    • v.32 no.1
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    • pp.154-161
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    • 2024
  • Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disorder that causes progressive paralysis. L-Citrulline is a nonessential neutral amino acid produced by L-arginine via nitric oxide synthase (NOS). According to previous studies, the pathogenesis of ALS entails glutamate toxicity, oxidative stress, protein misfolding, and neurofilament disruption. In addition, L-citrulline prevents neuronal cell death in brain ischemia; therefore, we investigated the change in the transport of L-citrulline under various pathological conditions in a cell line model of ALS. We examined the uptake of [14C]L-citrulline in wild-type (hSOD1wt/WT) and mutant NSC-34/ SOD1G93A (MT) cell lines. The cell viability was determined via MTT assay. A transport study was performed to determine the uptake of [14C]L-citrulline. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis was performed to determine the expression levels of rat large neutral amino acid transported 1 (rLAT1) in ALS cell lines. Nitric oxide (NO) assay was performed using Griess reagent. L-Citrulline had a restorative effect on glutamate induced cell death, and increased [14C]L-citrulline uptake and mRNA levels of the large neutral amino acid transporter (LAT1) in the glutamate-treated ALS disease model (MT). NO levels increased significantly when MT cells were pretreated with glutamate for 24 h and restored by co-treatment with L-citrulline. Co-treatment of MT cells with L-arginine, an NO donor, increased NO levels. NSC-34 cells exposed to high glucose conditions showed a significant increase in [14C]L-citrulline uptake and LAT1 mRNA expression levels, which were restored to normal levels upon co-treatment with unlabeled L-citrulline. In contrast, exposure of the MT cell line to tumor necrosis factor alpha, lipopolysaccharides, and hypertonic condition decreased the uptake significantly which was restored to the normal level by co-treating with unlabeled L-citrulline. L-Citrulline can restore NO levels and cellular uptake in ALS-affected cells with glutamate cytotoxicity, pro-inflammatory cytokines, or other pathological states, suggesting that L-citrulline supplementation in ALS may play a key role in providing neuroprotection.

Nitric Oxide Synthase Inhibitor Decreases NMDA-Induced Elevations of Extracellular Glutamate and Intracellular $Ca^{2+}$ Levels Via a cGMP-Independent Mechanism in Cerebellar Granule Neurons

  • Oh, Sei-Kwan;Yun, Bong-Sik;Ryoo, In-Ja;Patrick P.McCaslin;Yoo, Ick-Dong
    • Archives of Pharmacal Research
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    • v.22 no.1
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    • pp.48-54
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    • 1999
  • These studies were designed to examine the differential effect of nitric oxide (NO) and cGMP on glutamate neurotransmission. In primary cultures of rat cerebellar granule cells, the glutamate receptor agonist N-methyl-D-aspartate (NMDA) stimulates the elevation of intracellular calcium concentration ($[Ca^{2+}]_i$), the release of glutamate, the synthesis of NO and an increase of cGMP. Although NO has been shown to stimulate guanylyl cyclase, it is unclear yet whether NO alters the NMDA-induced glutamate release and ${[Ca^{2+}]}_i$ elevation. We showed that the NO synthase inhibitor, NG-monomethyl-L-arginine (NMMA), partially prevented the NMDA-induced release of glutamate and elevation of ${[Ca^{2+}]}_i$ and completely blocked the elevation of cGMP. These effects of NO on glutamate release and [Ca2+]i elevation were unlikely to be secondary to cGMP as the cGMP analogue, dibutyryl cGMP (dBcGMP), did not suppress the effects of NMDA. Rather, dBcGMP slightly augmented the NMDA-induced elevation of ${[Ca^{2+}]}_i$ with no change in the basal level of glutamate or ${[Ca^{2+}]}_i$. The extracellular NO scavenger hydroxocobalamine prevented the NMDA-induced release of glutamate providing indirect evidence that the effect of NO may act on the NMDA receptor. These results suggest that low concentration of NO has a role in maintaining the NMDA receptor activation in a cGMP-independent manner.

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Study on Production of Poly-γ-Glutamic Acid by Bacillus subtilis CH-10 (Bacillus subtilis CH-10에 의한 폴리감마글루탐산의 생산에 관한 연구)

  • Gu Na-Yeon;Kim Choon-Hee;Kim Byung-Woo;Nam Soo-Wan;Kwon Hyun-Ju;Kim Dong-Eun;Kim Young-Man;Jeon Sung-Jong
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.175-179
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    • 2006
  • A bacterium that produced a large amount of poly-$\gamma$-glutamate (PGA) was isolated from the compost and designated as Bacillus subtilis CH-10. The optimum temperature and pH for PGA production were at $37^{\circ}C$ and 7.5, respectively. The maximum amount of PGA production (18.84 mg/ml) was obtained when it was grown in a medium containing 3% L-glutamate and 5% sucrose at $37^{\circ}C$ with shaking. The result that the L-glutamate significantly induced PGA production indicates that it produces a PGA by the glutamate dependent manner. Some properties of the PGA obtained at different times of cultivation were investigated by SDS-PAGE and ninhydrin analysis. The PGA production was elongated along with cultivation time and maximum amount was achieved at 96 h. Average molecular weight of PGA was estimated to be 1100 kDa by FDNB method.

Studies on the Bacterial Production of L-Glutamate from Acetate Part I. Screening and Identification of L-Glutamate Producing Bacteria. (초산을 이용한 글루타민산의 발효생산에 관한 연구 제 1보 글루타민산 생산균주의 분리 및 동정)

  • 하덕모;노완섭
    • Microbiology and Biotechnology Letters
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    • v.2 no.2
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    • pp.103-109
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    • 1974
  • In the cource of the studies on L-glutamic acid production from acetic acid, 383 strains capable of assimilating acetate as sole source of carbon were isolated from 279 kinds of soil sample. Out of them, 5 strains which produced relatively larger amount of L-glutamate from acetate were selected and named Brevibacterium flavum nov. sp. D1005B, Corynebacterium glutamicum nov. sp. D1025A, Brevib. flavum nov. sp. D2209B, Coryneb. acetoacidophilum nov. sp. D2212B and Coryneb. acetoacidophilum nov. sp. D2349A respectively.

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Effects of Chronic Lead Exposure on Glutamate Release and Uptake in Cerebellar Cells of Rat Pups

  • Yi, Eun-Young;Lim, Dong-Koo
    • Archives of Pharmacal Research
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    • v.21 no.2
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    • pp.113-119
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    • 1998
  • Changes in the release and uptake of glutamate in cerebellar granule and glial cells of offspring of lead-exposed mothers were determined. In cultured cerebellar granule cells exposed to lead for 5 days, glutamate release was less influenced upon N-methyl-D-aspartate (NMDA) stimulation than that in the control. Although the NMDA-stimulated release of glutamate in cerebellar granule cells prepared from lead-exposed first generation pups was not different from that of the control group, the S-nitroso-N-acetylpenicillamine (SNAP)-stimulated release of glutamate in cerebellar granule cells obtained from lead-treated pups was less elevated than that in the control. Furthermore, in cerebellar granule cells obtained from lead-exposed second generations pups, glutamate release did not respond to both NMDA and SNAP stimulation. In cerebellar glial cells exposed to lead, the basal glutamate uptake was not changed. However, the L-trans-pyrollidine-2,4-dicarboxylic acid (PDC)-blocking effects was significantly reduced. In glial cells obtained from lead-exposed pups, the glutamate uptake was also less blocked by PDC than that in the control. Further decreases in PDC-blocking effects were observed in cerebellar glial cells obtained from lead-treated second generation pups compared to those from the control group. These results indicate that lead exposure induces the changes in the sensitivities of the glutamate release and uptake transporter. In addition, these results suggest that lead exposure might affect the intracellular signalling pathway and transmission in glutamatergic nervous system.

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