• Title/Summary/Keyword: Glucose-1-phosphate

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Crystallization and Preliminary X-ray diffraction Studies of Salmonella typhimurium Deoxy-Thymidine Diphosphate-D-Glucose-4, 6-Dehydratase (Salmonella typhimurium의 Deoxy-Thymidine Diphosphate-D-Glucose-4, 6-Dehydratase의 결정화와 X-선 회절에 관한 연구)

  • 최희욱;박교선
    • Korean Journal of Crystallography
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    • v.7 no.2
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    • pp.120-125
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    • 1996
  • Single crystals of deoxy-thymidine diphosphate-D-gluxose-4,6-dehydratase(abbreviated as dTDP-D-glucose dehydratase) from Escherichia coli Strain BL21 clone which harbors the gene of dTDP-D-glucose dehydratase in Salmonella typhimurium LT2 have been grown with and whithout substrates by sitting drop vapor diffusion at room temperature. The precipitating agent was 1.6 to 2.0 M Na, K phosphate buffer(pH 8.0). The crystals diffract to at least 2.5Å and belong to the hexagonal space group P61 with cell dimensions a=b=168.54Å, c=81.08Å. The asymmetric unit contains one dimer with a crystal volume per protein mass(VM) of 2.4Å3/Da and solvent content (Vsol) of 64% by volume.

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Determination of the Intracellular Concentrations of Metabolites in Escherichia coli Collected during the Exponential and Stationary Growth Phases using Liquid Chromatography-Mass Spectrometry

  • Park, Chang-Hun;Park, Chang-Hun;Lee, Youn-Jin;Lee, Sang-Yup;Oh, Han-Bin;Lee, Jin-Won
    • Bulletin of the Korean Chemical Society
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    • v.32 no.2
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    • pp.524-530
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    • 2011
  • In the present study, we demonstrate that SRM LC-MS/MS method developed by Luo et al. (ref. 10) can be successfully applied to the quantitative analysis of intracellular metabolites in E. coli that are collected at the exponential and stationary growth phases. A focus is given on measuring the changes in the concentrations of intracellular metabolites in batch cultures, which were induced during both the dynamically changing exponential and stationary growth phases. The following intracellular metabolites are quantified in the exponential and stationary phases of E. coli growth, using the SRM mode of a triple quadrupole mass spectrometer: glucose-1-phosphate, fructose-1,6-bisphosphate, phosphoenolpyruvate, pyruvate, acetyl-coenzyme A, 6-phosphogluconate, ribulose-5-phosphate, xylulose-5-phosphate, erythrose-4-phosphate. The determined intracellular metabolite concentration profiles are shown to be in a good agreement with the growth profiles of E. coli, which clearly indicates that SRM LC-MS/MS can be successfully used for following the metabolite changes induced at different growth stages.

Effect of Glucose on Listeria monocytogenes Survival under Sequential Sublethal Stresses of Gamma Irradiation and NaCl

  • Yoon, Yo-Han;Kim, Gyeong-Yeol;Nam, Min-Ji;Shim, Won-Bo;Seo, Eun-Kyoung;Kim, Jae-Hun;Lee, Ju-Woon;Byun, Myung-Woo;Chung, Duck-Hwa
    • Food Science and Biotechnology
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    • v.18 no.1
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    • pp.162-166
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    • 2009
  • This study evaluated glucose effect on Listeria monocytogenes survival under gamma irradiation and NaCl stress. L. monocytogenes in phosphate buffered saline (PBS) plus glucose (0-4%) was treated with gamma irradiation (0-0.5 kGy), and the samples were then exposed to NaCl (0-9%) in tryptic soy agar plus 0.6% yeast extract. $D_{10}$ and $t_{3D}$ values were determined, and a model for prediction of $D_{10}$ values was developed. Cell counts of L. monocytogenes reduced as irradiation dose increased, and L. monocytogenes in PBS (no glucose) was more sensitive to irradiation and NaCl compared to those in PBS (2 or 4% glucose). $D_{10}$ values were 0.07-0.1, 0.12-0.16, and 0.13-0.15 kGy for 0, 2, and 4% glucose, respectively. The $t_{3D}$ values were 0.22-0.3 (0% glucose), 0.35-0.48 (2% glucose), and 0.40-0.44 (4% glucose). A model performance was acceptable. These results indicate that glucose in foods would increase the resistance of L. monocytogenes to gamma irradiation and NaCl stress.

Membrane Free Stem Cell Extract from Adipose Tissue Enhances Glucose Uptake in 3T3-L1 Cells (무막줄기세포추출물의 3T3-L1 세포에서 포도당 흡수 촉진 효과)

  • Kim, Ji Hyun;Kim, Min Jeong;Park, Hye Sook;Kim, Young Sil;Cho, Eun Ju
    • Journal of Korean Medicine for Obesity Research
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    • v.19 no.2
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    • pp.89-96
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    • 2019
  • Objectives: We investigated whether membrane free stem cell extract from adipose tissue (MFSCE) has anti-diabetic effect. Methods: To determine glucose uptake effect of MFSCE, we carried out glucose uptake assay in 3T3-L1 adipocytes. The regulatory mechanisms of MFSCE on glucose uptake were examined by Western blot analysis. Results: When MFSCE was treated to adipocytes at the concentration of 0.5, 1, 2.5, and 5 ㎍/mL, 2-deoxyglucose-6-phosphate uptake was elevated approximately 1.8-fold compared to cells not treated with MFSCE. It indicated that MFSCE enhances glucose uptake in 3T3-L1 adipocytes. In addition, MFSCE reduced phosphorylation of insulin receptor substrate-1 at serine 307 and induced Akt and glucose transporter 4 protein expressions that were related to insulin signaling. Furthermore, MFSCE regulated adenosine monophosphate-activated protein kinase (AMPK) pathway by increases of increase phosphorylation of AMPK and acetyl-CoA carboxylase that were related to AMPK pathway. Conclusions: These results indicated that MFSCE promotes glucose uptake via modulation of insulin signaling and AMPK pathway. Therefore, MFSCE could be a promising agent for treatment of diabetes mellitus.

Glucose-6-phosphate dehydrogenase deficiency does not increase the susceptibility of sperm to oxidative stress induced by H2O2

  • Roshankhah, Shiva;Rostami-Far, Zahra;Shaveisi-Zadeh, Farhad;Movafagh, Abolfazl;Bakhtiari, Mitra;Shaveisi-Zadeh, Jila
    • Clinical and Experimental Reproductive Medicine
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    • v.43 no.4
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    • pp.193-198
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    • 2016
  • Objective: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common human enzyme defect. G6PD plays a key role in the pentose phosphate pathway, which is a major source of nicotinamide adenine dinucleotide phosphate (NADPH). NADPH provides the reducing equivalents for oxidation-reduction reductions involved in protecting against the toxicity of reactive oxygen species such as $H_2O_2$. We hypothesized that G6PD deficiency may reduce the amount of NADPH in sperms, thereby inhibiting the detoxification of $H_2O_2$, which could potentially affect their motility and viability, resulting in an increased susceptibility to infertility. Methods: Semen samples were obtained from four males with G6PD deficiency and eight healthy males as a control. In both groups, motile sperms were isolated from the seminal fluid and incubated with 0, 10, 20, 40, 60, 80, and $120{\mu}M$ concentrations of $H_2O_2$. After 1 hour incubation at $37^{\circ}C$, sperms were evaluated for motility and viability. Results: Incubation of sperms with 10 and $20{\mu}M\;H_2O_2$ led to very little decrease in motility and viability, but motility decreased notably in both groups in 40, 60, and $80{\mu}M\;H_2O_2$, and viability decreased in both groups in 40, 60, 80, and $120{\mu}M\;H_2O_2$. However, no statistically significant differences were found between the G6PD-deficient group and controls. Conclusion: G6PD deficiency does not increase the susceptibility of sperm to oxidative stress induced by $H_2O_2$, and the reducing equivalents necessary for protection against $H_2O_2$ are most likely produced by other pathways. Therefore, G6PD deficiency cannot be considered as major risk factor for male infertility.

Organic acid production and phosphate solubilization by Enterobacter intermedium 60-2G (Enterobacter intermedium 60- 2G의 유기산 생성과 불용성인의 가용화)

  • Kim, Kil-Yong;Hwangbo, Hoon;Kim, Yong-Woong;Kim, Hyo-Jeong;Park, Keun-Hyung;Kim, Young-Cheol;Seong, Ki-Young
    • Korean Journal of Soil Science and Fertilizer
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    • v.35 no.1
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    • pp.59-67
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    • 2002
  • A phosphate solubilizing bacterium. strain 60-2G, possessing a strong ability to solubilize insoluble phosphate was isolated from the rhizosphere of grass. On the basis of GC-FAME profile, carbon utilization pattern, and the DNA sequence of a conserved partial 16S rRNA gene, the 60-2G was identified as Enterobacter intermedium. The analysis by HPLC revealed that the strain 60-2G produced mainly gluconic and 2-ketogluconic acids with small amounts of lactic acid in broth culture medium containing hydroxyapatite. During the incubation period of the strain 60-2G in broth culture, pH of the medium decreased upto 3.8 while the soluble phosphate concentration increased. The reversed correlation between pH and soluble phosphate concentration indicated that the solubility of P was due to the produced organic acids. The sequence homology of the deduced amino acids suggested that E. intermedium 60-2G synthesized PQQ which is essential for the oxidation of glucose by glucose dehydrogenase.

Isolation and Characterization of Hydrogen Producing Bacterium (수소생산균 Enterobacter cloacae YJ -1의 분리 및 특성)

  • 이기석;강창민;정선용
    • KSBB Journal
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    • v.18 no.2
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    • pp.149-154
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    • 2003
  • The hydrogen-producing bacterium was isolated from fresh water and identified as Enterobacter cloacae. The isolated was named Enterobacter cloacae YJ-1. In batch culture, The optimum cultivation temperature and pH of strain YJ-1 was 35℃ and 7.5, respectively. All of the added glucose was consumed completely during fermentation even though pH was not controlled. Amount of hydrogen produced on each condition of 2% glucose, 4% sucrose and 5% fructose was 950, 1000 and 948 mL/L, respectively and resulted in increasing hydrogen production approximately 2.5-times more than controlled condition. The maximum hydrogen production was obtained when 50 mM phosphate was added. In repeated-batch culture, hydrogen gas of 1920 mL/L was totally produced for 48. The maximum hydrogen was produced on the condition of 0.5% yeast extract, but the production amount was not changed on the condition of over 0.5%. Most of the organic acids produced during the fermentation were formic and acetic acid, and propionic acid was moiety also generated.

Myo-Inositol Synthesis in the Milk of Lactating Rats (쥐 우유중의 Myo-Inositol 생성에 관한 연구)

  • Byun, Si-Myung
    • Applied Biological Chemistry
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    • v.19 no.3
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    • pp.121-129
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    • 1976
  • A high concentration of myo-Inositol in rat's milk was observed (61-91mg. of myo-Inositol per 100g of milk) by gas-liquid chromatographic method, using a 3% SE-52 column. Feeding experiments showed that approximately 85% of myo-Inositol in milk was from dietary origin: the rest was considered to be synthesized by 1L-myo-Inositol-1-phosphate lyase. Results suggested that the biosynthesis was not sufficiently high to permit the maintenance of its myo-Inositol level in milk. However, study $using(^{14}C)-glucose$ injection into lactating female rats confirmed biosynthesis of myo-Inositol from glucose in mammary gland. This biosynthesis reached a maximum within an hour after $(^{14}C)-glucose$ injection intraperitoneally as lactose biosynthesis did. Study using $(^3H)-myo-Inositol$ confirmed that most of the myo-Inositol in milk was transported from blood plasma myo-Inositol against a concentration gradient. About four hours after the beginning of the injection of $(^{14}C)-glucose$, the specific radioactivity of myo-Inositol in milk was 8% of that of glucose in the blood. When $(^3H)-myo-Inositol$ was injected, the specific radioactivity of myo-Inositol in milk was about 26% of that of blood six hours after injection.

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Antidiabetic and Antioxidative Effects of Cibotium barometz in Streptozotocin-Induced Diabetic Rats (구척 에탄올 추출물이 Streptozotocin 유발 당뇨 흰쥐에 대한 항당뇨 및 항산화 작용에 미치는 효과)

  • Kim, Ok-Kyung
    • Journal of the Korean Applied Science and Technology
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    • v.32 no.3
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    • pp.488-496
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    • 2015
  • This study was done to investigate the antidiabetic and antioxidant effects of Cibotium barometz in Streptozotocin (STZ)-induced diabetic rats. Diabetes was induced by intravenous injection of STZ at a dose 45mg/kg.b.w. dissolved in citrate buffer(pH4.5). The ethanol extract of Cibotium barometz was orally administrated once a day for 7 days. The contents of serum glucose, triglyceride(TG) and total cholesterol were significantly decreased(p<0.05) in Cibotium barometz treated group compared to the those of STZ-control group, The content of glutathione(GSH) and activity of gluthathione-s-transferase(GST) were significantly increased (P<0.05) in Cibotium barometz treated group compared to the those of STZ-control group. and activityes of catalase(CAT) and glutathione peroxidae(GSH-Px) were signiicantly decreased (P<0.05) in Cibotium barometz treated group compared to the those of STZ-control group. Also the content of hepatic glycogen and activities of glucose-phosphate dehydrogenase(G-6-PDH)and glucokinase(GK) were significamtly increased(p<0.05), but activity of glucose-6-phosphatase (G-6-Pase) was significamtly decreased (p<0.05) in Cibotium barometz treated group compared to the those of STZ-control group. These results indicated that ethanol extract of Cibotium barometz would have antidiabetic and antioxidant effects in STZ-induced diabetic rats.

Role of Phospholipase $A_2$ in Oxidant-induced Alteration in Phosphate Transport in Primary Cultured Rabbit Renal Proximal Tubule Cells

  • Park, Kwon-Moo;Ko, Sun-Hee;Woo, Jae-Suk;Jung, Jin-Sup;Lee, Sang-Ho;Kim, Yong-Keun
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.5
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    • pp.601-609
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    • 1998
  • The present study was undertaken to examine the role of phospholipase $A_2\;(PLA_2)$ in oxidant-induced inhibition of phosphate transport in primary cultured rabbit renal proximal tubule cells. Uptakes of phosphate and glucose were dose-dependently inhibited by an oxidant t-butylhydroperoxide (tBHP), and the significant inhibition appeared at 0.025 mM of tBHP, whereas tBHP-induced alterations in lipid peroxidation and cell viability were seen at 0.5 mM. tBHP stimulated arachidonic acid (AA) release in a dose-dependent fashion. A $PLA_2$ inhibitor mepacrine prevented tBHP-induced AA release, but it did not alter the inhibition of phosphate uptake and the decrease in cell viability induced by tBHP. tBHP-induced inhibition of phosphate transport was not affected by a PKC inhibitor, staurosporine. tBHP at 0.1 mM did not produce the inhibition of $Na^+-K^+-ATPase$ activity in microsomal fraction, although it significantly inhibited at 1.0 mM. These results suggest that tBHP can inhibit phosphate uptake through a mechanism independent of $PLA_2$ activation, irreversible cell injury, and lipid peroxidation in primary cultured rabbit renal proximal tubular cells.

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