• Title/Summary/Keyword: L-glutamate

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Protective Effect of Sanguisorba officinalis L. Root on Amyloid ${\beta}$ Protein (25-35)-induced Neuronal Cell Damage in Cultured Rat Cortical Neuron

  • Ban, Ju-Yeon;Cho, Soon-Ock;Jeon, So-Young;Song, Kyung-Sik;Bae, Ki-Hwan;Seong, Yeon-Hee
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.5
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    • pp.219-226
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    • 2005
  • Sanguisorbae radix (SR) from Sanguisorba officinalis L. (Losaceae) is widely used in Korea and China due to its various pharmacological activity. The present study aims to investigate the effect of the methanol extract of SR on amyloid ${\beta}$ Protein(25-35) $(A{\beta}\;(25-35))$, a synthetic 25-35 amyloid peptide, -induced neurotoxicity using cultured rat cortical neurons. SR, over a concentration range of $10-50\;{\mu}g/ml$, inhibited the $A{\beta}$ (25-35) $(10\;{\mu}M)-induced$ neuronal cell death, as assessed by a 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and the number of apoptotic nuclei, evidenced by Hoechst 33342 staining. Pretreatment of SR $(50\;{\mu}g/ml)$ inhibited $10\;{\mu}M\;A{\beta}$ (25-35)-induced} elevation of cytosolic calcium concentration $([Ca^{2+}]c)$, which was measured by a fluorescent dye, fluo-4 AM. SR $(10\;and\;50\;{\mu}g/ml)$ inhibited glutamate release into medium induced by $10\;{\mu}M\;A{\beta}(25-35)$, which was measured by HPLC, and generation of reactive oxygen species. These results suggest that SR prevents $A{\beta}$ (25-35)-induced neuronal cell damage in vitro.

Effect of Medicinal Plant Extracts on Alcohol Metabolism in Rat Liver

  • Lee, Seung-Eun;Bang, Jin-Ki;An, Tae-Jin;Yu, Young-Ju;Chung, Hae-Gon;Kim, Geum-Suk;Seong, Nak-Sul
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.2
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    • pp.113-117
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    • 2004
  • The experiment was conducted to evaluate the effects of medicinal plants on ethanol-metabolism. Sprague Dawley rats divided into 6 groups (n=8), fed with 10% ethanol and diets supplemented with each 1% of four plant extracts, ${\alpha}-tocopherol$ (as positive control) and fiber (as negative control) for 4 weeks. Group supplemented with plant extract of Ulmus davidiana showed the most high value (322 nM NADH/min/mg protein) in alcohol dehydrogenase (ADH) activity among the experimented groups $(144{\sim}312\;nM\;NADH/min/mg\;protein)$ at p<0.05. Groups fed with Lagerstroemia indica and Zelkova serrata extract-supplemented diets indicated high activity in aldehyde dehydrogenase (ALDH, 16.7 & 12.3 M NADH/min/mg protein), which were comparatively lower than 20.1 M NADH/min/mg protein of ${\alpha}-tocopherol$ fed group. All of the groups fed with plant extracts indicated very low GPT activities $(13.9{\sim}17.3\;IU/l)$ compared to those (146.1 & 128.6 IU/l) fed with ${\alpha}-tocopherol$ and fiber at p<0.05. From these results, it is suggested that Lagerstroemia indica have a potent ethanol-metabolizing activity.

Studies on Microbial Penicillin Amidase ( I ) Optimization of the Enzyme Production from Escherichia coli (미생물 페니실린 아미다제에 관한 연구 (I) E. coli로부터 효소생산 조건의 최적화)

  • Kim, Bong-Hee;Seong, Baik-Lin;Mheen, Tae-Iek;Moon H. Ban
    • Microbiology and Biotechnology Letters
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    • v.9 no.1
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    • pp.29-34
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    • 1981
  • To maximize the production of penicillin amidase from Estherichia coli (ATCC 9637), the media composition and several factors affecting the engyme production during fermentation were studied. The optimal media composition was found to be; 3.5% tryptone, 1.5% monosodium glutamate and 0.5% yeast extract. The addition of 0.15% phenylacetic acid as an enzyme inducer at the initial stage of cultivation increased the engyme productivity about 5 fold. It was found that the engyme activity reached maximum within 16hr of cultivation. The maximum production of the enzyme obtained was about 102.5 units/l broth under the optimized condition. The enzyme production was markedly increased by the optimization as compared with those previously reported.

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Bisphenol A Disturbs Intracellular Calcium Homeostasis and its Relationship with Cytotoxicity (Bisphenol A에 의한 신경계 세포의 칼슘 항상성 교란 및 세포독성에 미치는 영향)

  • Lee Yoot Mo;Lee Sang Min;Son Dong Ju;Lee Sun Young;;Nam Sang Yun;Kim Dae Joong;Yun Young Won;Yoo Hwan Soo;Oh Ki Wan;Kim Tae Seong;Han Soon Young;Hong Jin Tae
    • Toxicological Research
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    • v.20 no.3
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    • pp.241-250
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    • 2004
  • We previously found that bisphenol A (BPA) caused neurotoxic behavioral alteration. Since disturbance of calcium homeostasis is an implicated contributor in the neurotoxic mechanism of environmental toxicants, we investigated whether BPA alters calcium homeostasis. Unlike other neurotoxic agents which cause increase of intracellular calcium level, BPA decreased $[Ca^{2+}]_i$ dose-dependently in PC12 cells and cortical neuronal cells regardless of the calcium existence in buffer. BPA at greater concentrations than 100 $\mu\textrm{M}$ reduced cell viability significantly in both types of cells. BPA also suppressed L-glutamate (L-type channel activator, 30 mM) and trifluoperazine (calmodulin antagonist, 30 $\mu\textrm{M}$)-induced increase of $[Ca^{2+}]_i$. BPA further lowered caffeine (RYR activator, 100 $\mu\textrm{M}$)-decreased $[Ca^{2+}]_i$, but did not alter dantrolene (RYR inhibitor, 100 $\mu\textrm{M}$), heparin (IP3 inhibitor, 200 units/ml) and xestospongin C (IP3 inhibitor, 5 $\mu\textrm{M}$)-decreased $[Ca^{2+}]_i$. Cell viability was not directly related to intracellular calcium change by bisphenol A that alternation of intracellular calcium may not be a direct causal factor of BPA-induced neuronal cell death.

Expression and Purification of a Functional Recombinant Aspartate Aminotransferase (AST) from Escherichia coli

  • Zou, Lihui;Zhao, Haijian;Wang, Daguang;Wang, Meng;Zhang, Chuanbao;Xiao, Fei
    • Journal of Microbiology and Biotechnology
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    • v.24 no.7
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    • pp.998-1003
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    • 2014
  • Aspartate aminotransferase (AST; E.C. 2.6.1.1), a vitamin B6-dependent enzyme, preferentially promotes the mutual transformation of aspartate and ${\alpha}$-ketoglutarate to oxaloacetate and glutamate. It plays a key role in amino acid metabolism and has been widely recommended as a biomarker of liver and heart damage. Our study aimed to evaluate the extensive preparation of AST and its application in quality control in clinical laboratories. We describe a scheme to express and purify the 6His-AST fusion protein. An optimized sequence coding AST was synthesized and transformed into Escherichia coli BL21 (DE3) strain for protein expression. Ideally, the fusion protein has a volumetric productivity achieving 900 mg/l cultures. After affinity chromatography, the enzyme activity of purified AST reached 150,000 U/L. Commutability assessment between the engineered AST and standard AST from Roche suggested that the engineered AST was the better candidate for the reference material. Moreover, the AST showed high stability during long-term storage at $-20^{\circ}C$. In conclusion, the highly soluble 6His-tagged AST can become a convenient tool for supplying a much better and cheaper standard or reference material for the clinical laboratory.

Changes in Physicochemical and Sensory Properties of Hizikia fusiforme Water Extract by Fermentation of Lactic Acid Bacteria (유산균 발효에 의한 톳(Hizikia fusiforme) 추출액의 이화학적 및 관능적 특성 변화)

  • Song, Ho-Su;Kim, Hong-Kil;Min, Hye-Ok;Choi, Jong-Duck;Kim, Young-Mog
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.44 no.2
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    • pp.104-110
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    • 2011
  • This study was conducted to improve the food functional and sensory food quality of Hizikia fusiforme by the fermentation of lactic acid bacteria. Seven strains of lactic acid bacteria isolated from traditional Korean fermented food were inoculated and cultivated in H. fusiforme water extract. Among them, Lactobacillus brevis LB-20, isolated from Kimchi, was selected for further study by considering the results of bacterial growth, DPPH radical scavenging activity, and sensory evaluation. No significant differences in proximate compositions (moisture, crude protein, crude fat, and crude ash) were observed by the fermentation of L. brevis LB-20. The most dramatical change was the conversion from glutamate to ${\gamma}$-aminobutyric acid (GABA) in H. fusiforme water extract fermented by L. brevis LB-20. The GABA content increased approximately 60-fold after 48 hr of fermentation. The bacterial fermentation also resulted in low-molecularization of the extract. The particle size of the fermented extract became approximately 4-fold smaller than that of the law extract. In addition, the analysis of volatile flavor compounds using GC/MS revealed that the bacterial fermentation dramatically removed off-flavors such as acetaldehyde, haxanal, diallyl disulphide and 1-penten-2-ol in the H. fusiforme extract.

Processing and Characteristics of Canned Seasoned Sea Mussel (조미 홍합 통조림의 제조 및 특성)

  • Park, Tae-Ho;Noe, Yu-Ni;Lee, In-Seok;Kwon, Soon-Jae;Yoon, Ho-Dong;Kong, Cheung-Sik;Nam, Dong-Bae;Oh, Kwang-Soo;Kim, Jeong-Gyun
    • Journal of Fisheries and Marine Sciences Education
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    • v.24 no.6
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    • pp.820-832
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    • 2012
  • This study was investigated to obtain basic data which can be applied to processing of canned seasoned sea mussel. Shell was washed and steamed for 10 min before shucking. Sea mussel meat was seasoned with mixed seasoning sauce(soy sauce 23%, monosodiun glutamate 2%, sorbitol 2%, sesame oil 1%, vinegar 2%, starch syrup 15%, water 55%) for 30 min The seasoned sea mussel 60 g was vacuum packed in RR-90 can and fill with seasoning sauce 30 mL and grape seed oil 30 mL respectively, and then there was sterilized for various Fo values(Fo 8~12 min) in a steam system retort at $121^{\circ}C$. pH, VBN, amino-N, total amino acid, free amino acid, color value, texture profile, TBA value, mineral content, sensory evaluation and viable cells count of the canned seasoned sea mussels sterilized with various conditions(Fo 8~12 min) were measured. The same experimental items were also measured during storage. There was no remarkable difference between sterilized conditions and sensual characteristics. The results showed that the product of filled with grape seed oil sterilized at Fo 8 min was the most desirable.

Growth Inhibition of Escherichia coli during Heterologous Expression of Bacillus subtilis Glutamyl-tRNA Synthetase that Catalyzes the Formation of Mischarged Glutamyl-$tRNA_{l}$$^{Gln}$

  • Baick, Ji-Won;Yoon, Jang-Ho;Suk Namgoong;Dieter Soll;Kim, Sung-Il;Eom, Soo-Hyun;Hong, Kwang-Won
    • Journal of Microbiology
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    • v.42 no.2
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    • pp.111-116
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    • 2004
  • It is known that Bacillus subtilis glutamyl-tRNA synthetase (GluRS) mischarges E. coli $tRNA_{1}$$^{Gln}$ with glutamate in vitro. It has also been established that the expression of B. subtilis GluRS in Escherichia coli results in the death of the host cell. To ascertain whether E. coli growth inhibition caused by B. subtilis GluRS synthesis is a consequence of Glu-$tRNA_{1}$$^{Gln}$ formation, we constructed an in vivo test system, in which B. subtilis GluRS gene expression is controlled by IPTG. Such a system permits the investigation of factors affecting E. coli growth. Expression of E. coli glutaminyl-tRNA synthetase (GlnRS) also amelio-rated growth inhibition, presumably by competitively preventing $tRNA_{1}$$^{Gln}$ misacylation. However, when amounts of up to 10 mM L-glutamine, the cognate amino acid for acylation of $tRNA_{1}$$^{Gln}$, were added to the growth medium, cell growth was unaffected. Overexpression of the B. subtilis gatCAB gene encoding Glu-$tRNA^{Gln}$ amidotransferase (Glu-AdT) rescued cells from toxic effects caused by the formation of the mis-charging GluRS. This result indicates that B. subtilis Glu-AdT recognizes the mischarged E. coli Glu-$tRNA_{1}$$^{Gln}$, and converts it to the cognate Gln-$tRNA_{1}$$^{Gln}$ species. B. subtilis GluRS-dependent Glu-$tRNA_{1}$$^{Gln}$ formation may cause growth inhibition in the transformed E. coli strain, possibly due to abnormal protein synthesis.

Vascular Endothelial Growth Factor May Be Involved in the Behavioral Changes of Progeny Rats after Exposure to Ceftriaxone Sodium during Pregnancy

  • Yang, Xin;Tang, Ting;Li, Mengchun;Chen, Jie;Li, Tingyu;Dai, Ying;Cheng, Qian
    • Journal of Microbiology and Biotechnology
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    • v.32 no.6
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    • pp.699-708
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    • 2022
  • Antibiotic exposure during pregnancy have an adversely effects on offspring behavior and development. However, its mechanism is still poorly understood. To uncover this, we added ceftriaxone sodium to the drinking water of rats during pregnancy and conducted three-chamber sociability test, open-field test, and Morris water maze test in 3- and 6-week-old offspring. The antibiotic group offspring showed lower sociability and spatial learning and memory than control. To determine the role of the gut microbiota and their metabolites in the changes in offspring behavior, fecal samples of 6-week-old offspring rats were sequenced. The composition of dominant gut microbial taxa differed between the control and antibiotic groups. KEGG pathway analysis showed that S24-7 exerted its effects through the metabolic pathways including mineral absorption, protein digestion and absorption, Valine, leucine, and isoleucine biosynthesis. Correlation analysis showed that S24-7 abundance was negatively correlated with the level of VEGF, and metabolites associated with S24-7-including 3-aminobutanoic acid, dacarbazine, L-leucine, 3-ketosphinganine, 1-methylnicotinamide, and N-acetyl-L-glutamate-were also significantly correlated with VEGF levels. The findings suggest that antibiotic exposure during pregnancy, specifically ceftriaxone sodium, will adversely affects the behavior of offspring rats due to the imbalance of gut microbiota, especially S24-7, via VEGF and various metabolic pathways.

Ammonium Excess Promotes Proline Synthesis but Inhibits Glutathione Synthesis in Oilseed Rape (Brassica napus L.)

  • Hyunjae Lee;Seon-Hye Baek;Tae-Hwan Kim
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.43 no.2
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    • pp.109-115
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    • 2023
  • Ammonium (NH4+) serves as a nitrogen source, but its elevated levels can hinder plant growth and production. Excess NH4+ with α-ketoglutarate is assimilated into glutamate, a precursor of proline and glutathione (GSH). This study aimed to investigate the effects of excessive NH4+ on the regulation of proline and GSH synthesis. Detached leaves from oilseed rape (Brassica napus L.) were fed with 0, 50, 100, 500, and 1000 mM NH4Cl for 16 h. As the NH4+ concentrations increased, the leaves exhibited progressive wilting and yellowing. Furthermore, total carotenoid and chlorophyll concentrations declined in response to all NH4+ treatments, with the lowest levels observed in 1000 mM NH4+ treatment. Hydrogen peroxide (H2O2) concentration showed a minor increase at low NH4+ concentration (50 and 100 mM) treatments but a significant increase at high NH4+ (500 and 1000 mM), which was consistent with the localization of H2O2. Amino acid concentrations increased with increasing in NH4+ concentration, while the protein concentration displayed the opposite trend. Proline and cysteine concentrations exhibited a gradual increase in response to increasing NH4+ concentrations. However, GSH concentrations rose only in the 50 mM NH4+ treatment and decreased in the 500 and 1000 mM NH4+ treatments. These results indicate that excessive NH4+ is primarily assimilated into proline, while GSH synthesis is adversely affected.