• Title/Summary/Keyword: Phosphate stress

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Isoforms of Glucose 6-Phosphate Dehydrogenase in Deinococcus radiophilus

  • Sung, Ji-Youn;Lee, Young-Nam
    • Journal of Microbiology
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    • v.45 no.4
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    • pp.318-325
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    • 2007
  • Glucose 6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) in Deinococcus radiophilus, an extraordinarily UV-resistant bacterium, was investigated to gain insight into its resistance as it was shown to be involved in a scavenging system of superoxide $(O_2^{-1})$ and peroxide $(O_2^{-2})$ generated by UV and oxidative stresses. D. radiophilus possesses two G6PDH isoforms: G6PDH-1 and G6PDH-2, both showing dual coenzyme specificity for NAD and NADP. Both enzymes were detected throughout the growth phase; however, the substantial increase in G6PDH-1 observed at stationary phase or as the results of external oxidative stress indicates that this enzyme is inducible under stressful environmental conditions. The G6PDH-1 and G6PDH-2 were purified 122- and 44-fold (using NADP as cofactor), respectively. The purified G6PDH-1 and G6PDH-2 had the specific activity of 2,890 and 1,033 U/mg protein (using NADP as cofactor) and 3,078 and 1,076 U/mg protein (using NAD as cofactor), respectively. The isoforms also evidenced distinct structures; G6PDH-1 was a tetramer of 35 kDa subunits, whereas G6PDH-2 was a dimer of 60kDa subunits. The pIs of G6PDH-1 and G6PDH-2 were 6.4 and 5.7, respectively. Both G6PDH-1 and G6PDH-2 were inhibited by both ATP and oleic acid, but G6PDH-1 was found to be more susceptible to oleic acid than G6PDH-2. The profound inhibition of both enzymes by ${\beta}-naphthoquinone-4-sulfonic$ acid suggests the involvement of lysine at their active sites. $Cu^{2+}$ was a potent inhibitor to G6PDH-2, but a lesser degree to G6PDH-1. Both G6PDH-1 and G6PDH-2 showed an optimum activity at pH 8.0 and $30^{\circ}C$.

Fed-batch Cultivation of Escherichia coli YK537 (pAET-8) for Production of phoA Promoter-controlled Human Epidermal Growth Factor

  • Wang Yonggang;Du Peng;Gan Renbao;Li Zhimin;Ye Qin
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.2
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    • pp.149-154
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    • 2005
  • Secretion of the expressed heterologous proteins can reduce the stress to the host cells and is beneficial to their recovery and purification. In this study, fed-batch cultures of Escherichia coli YK537 (pAET-8) were conducted in a 5-L fermentor for the secretory production of human epidermal growth factor (hEGF) whose expression was under the control of alkaline phosphatase promoter. The effects of feeding of glucose and complex nitrogen sources on hEGF production were investigated. When the fed-batch culture was conducted in a chemically de-fined medium, the cell density was 9.68 g/L and the secreted hEGF was 44.7 mg/L in a period of 60 h. When a complex medium was used and glucose was added in pH-stat mode, the secreted hEGF was improved to 345 mg/L. When the culture was fed with glucose at a constant specific rate of $0.25\;gg^{-1}h^{-1}$, hEGF reached 514 mg/L. The effects of adding a solution containing yeast extract and tryptone were further studied. Different rate of the nitrogen source feeding resulted in different levels of phosphate and acetic acid formation, thus affected hEGF expression. At the optimal feeding rate, hEGF production achieved 686 mg/L.

Dexamethasone enhances glucose uptake by SGLT1 and GLUT1 and boosts ATP generation through the PPP-TCA cycle in bovine neutrophils

  • Wang, Xinbo;Tang, Mingyu;Zhang, Yuming;Li, Yansong;Mao, Jingdong;Deng, Qinghua;Li, Shusen;Jia, Zhenwei;Du, Liyin
    • Journal of Veterinary Science
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    • v.23 no.5
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    • pp.76.1-76.14
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    • 2022
  • Background: Clinical dexamethasone (DEX) treatment or stress in bovines results in extensive physiological changes with prominent hyperglycemia and neutrophils dysfunction. Objectives: To elucidate the effects of DEX treatment in vivo on cellular energy status and the underlying mechanism in circulating neutrophils. Methods: We selected eight-month-old male bovines and injected DEX for 3 consecutive days (1 time/d). The levels of glucose, total protein (TP), total cholesterol (TC), and the proinflammatory cytokines interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α in blood were examined, and we then detected glycogen and adenosine triphosphate (ATP) content, phosphofructosekinase-1 (PFK1) and glucose-6-phosphate dehydrogenase (G6PDH) activity, glucose transporter (GLUT)1, GLUT4, sodium/glucose cotransporter (SGLT)1 and citrate synthase (CS) protein expression and autophagy levels in circulating neutrophils. Results: DEX injection markedly increased blood glucose, TP and TC levels, the Ca2+/P5+ ratio and the neutrophil/lymphocyte ratio and significantly decreased blood IL-1β, IL-6 and TNF-α levels. Particularly in neutrophils, DEX injection inhibited p65-NFκB activation and elevated glycogen and ATP contents and SGLT1, GLUT1 and GR expression while inhibiting PFK1 activity, enhancing G6PDH activity and CS expression and lowering cell autophagy levels. Conclusions: DEX induced neutrophils glucose uptake by enhancing SGLT1 and GLUT1 expression and the transformation of energy metabolism from glycolysis to pentose phosphate pathway (PPP)-tricarboxylic acid (TCA) cycle. This finding gives us a new perspective on deeper understanding of clinical anti-inflammatory effects of DEX on bovine.

Gravitropism in the Salt-Stressed Primary Root of Maize (Zea mays) (염분 스트레스에 노출된 옥수수(Zea mays) 뿌리의 굴중성 반응)

  • Han, Du-Yeol;Lee, Young-Na;Kim, Yeo-Jae;Park, Woong-June
    • Journal of Life Science
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    • v.18 no.8
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    • pp.1164-1168
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    • 2008
  • We investigated gravitropic responses in the primary root of maize (Zea mays) seedlings which were exposed to salt stress. The maize roots salt-stressed with higher than 100 mM NaCl or KCl started to reveal enhanced gravitropic curvature after 2 hours form the gravi-stimulation. Such a promotion was not caused by sodium phosphate, but invoked by potassium phosphate, indicating the active component is $K^{+}$. Because NaCl increased gravitropic curvature, despite that $Na^{+}$ did not played any role, we evaluated the role for $Cl^{-}$ by comparing the effects of $MgCl_2$ and $MgSO_4$. The enhancement of the curvature only with $MgCl_2$ revealed that $Cl^{-}$ played a role in the gravi-response, indicating the involvement of anion channels. These results suggest that both of $K^{+}$ and $Cl^{-}$ play roles in the regulation of osmosis that is required for cell expansion in gravitropism as well as in nyctinasty and stomatal opening.

Potential involvement of Drosophila flightless-1 in carbohydrate metabolism

  • Park, Jung-Eun;Jang, Jinho;Lee, Eun Ji;Kim, Su Jung;Yoo, Hyun Ju;Lee, Semin;Kang, Min-Ji
    • BMB Reports
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    • v.51 no.9
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    • pp.462-467
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    • 2018
  • A previous study of ours indicated that Drosophila flightless-1 controls lipid metabolism, and that there is an accumulation of triglycerides in flightless-1 (fliI)-mutant flies, where this mutation triggers metabolic stress and an obesity phenotype. Here, with the aim of characterizing the function of FliI in metabolism, we analyzed the levels of gene expression and metabolites in fliI-mutant flies. The levels of enzymes related to glycolysis, lipogenesis, and the pentose phosphate pathway increased in fliI mutants; this result is consistent with the levels of metabolites corresponding to a metabolic pathway. Moreover, high-throughput RNA sequencing revealed that Drosophila FliI regulates the expression of genes related to biological processes such as chromosome organization, carbohydrate metabolism, and immune reactions. These results showed that Drosophila FliI regulates the expression of metabolic genes, and that dysregulation of the transcription controlled by FliI gives rise to metabolic stress and problems in the development and physiology of Drosophila.

Screening Differential Expressions of Defense-related Responses in Cold-treated 'Kyoho' and 'Campbell Early' Grapevines

  • Ahn, Soon Young;Kim, Seon Ae;Han, Jae Hyun;Kim, Seung Heui;Yun, Hae Keun
    • Horticultural Science & Technology
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    • v.31 no.3
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    • pp.275-281
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    • 2013
  • Low temperature is one of the major environmental factors that affect productivity including reduced growth and budding of vines, and changes of metabolic processes in grape (Vitis spp.). To screen the specific expression of abiotic stress-related genes against cold treatment in 'Kyoho' and 'Campbell Early' grapevines, expression of various defense-related genes was investigated by RT-PCR and real-time PCR. Among the 67 genes analyzed by RT-PCR and real-time PCR, 17 and 16 types of cDNA were up-regulated, while 5 and 6 types were down-regulated in cold-treated 'Kyoho' and 'Campbell Early' grapevines, respectively. Genes encoding carotene (Cart3564 and Cart4472), chalcone isomerase (CHI), cytochrome P450 (CYP), flavonol synthase (FLS), endo-${\beta}$-glucanase precursor (Glu), glutathione peroxidase (GPX), glutathione-S-transferase (GST), leucine-rich repeats (LRR), manganese superoxide dismutase (Mn-SOD), phenylalanine ammonia lyase (PAL), polygalacturonase-inhibiting protein (PGIP), proline rich protein 2 (PRP2), small heat shock protein (sHSP), temperature induced lipocalin (TIL), and thaumatin-like protein (TLP) were up-regulated, while those encoding CBF like transcription factor (CBF1), chitinase-like protein (CLP), cold induced protein (CIP), glycerol-3-phosphate acyltransferase (GPAT), and mitogen-activated protein kinase (MAPK) were down-regulated by low temperature treatment in both in 'Kyoho' and 'Campbell Early'.

Isolation and Characterization of Bifidobacterium longum subsp. longum BCBR-583 for Probiotic Applications in Fermented Foods

  • Yi, Da Hye;Kim, You-Tae;Kim, Chul-Hong;Shin, Young-Sup;Lee, Ju-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.28 no.11
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    • pp.1846-1849
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    • 2018
  • Recent human gut microbiome studies have supported that the genus Bifidobacterium is one of the most beneficial bacteria for human intestinal health. To develop a new probiotic strain for functional food applications, fourteen fecal samples were collected from healthy Koreans and the strain BCBR-583 was newly selected and isolated from a 25-year-old Korean woman's fecal sample using the selective medium for Bifidobacterium. Subsequent fructose-6-phosphate phosphoketolase (F6PPK) test and 16S rRNA gene sequencing analysis of the strain BCBR-583 confirmed that it belongs to B. longum subsp. longum. The stress resistance tests showed that it has oxygen and heat tolerance activities (5- and 3.9-fold increase for 24 h at 60 and 120 rpm, respectively; $78.61{\pm}6.67%$ survival rate at $45^{\circ}C$ for 24 h). In addition, gut environment adaptation tests revealed that this strain may be well-adapted in the gut habitat, with gastric acid/bile salt resistance ($85.79{\pm}1.53%$, survival rate under 6 h treatments of gastric acid and bile salt) and mucin adhesion ($73.72{\pm}7.36%$). Furthermore, additional tests including cholesterol lowering assay showed that it can reduce $86.31{\pm}1.85%$ of cholesterol. Based on these results, B. longum BCBR-583 has various stress resistance for survival during food processing and environmental adaptation activities for dominant survival in the gut, suggesting that it could be a good candidate for fermented food applications as a new probiotic strain.

Effect of Kinematic Motion on Changes in Coefficients of Friction of Porcine Knee Joint Cartilage (기구학적 운동이 돼지 무릎 관절연골의 마찰계수 변화에 미치는 영향)

  • Kim, Hwan;Kim, ChoongYeon;Lee, KwonYong;Kim, DaeJoon;Kim, DoHyung
    • Tribology and Lubricants
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    • v.29 no.1
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    • pp.46-50
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    • 2013
  • In this study, the frictional behaviors of articular cartilage against a Co-Cr alloy in two types of kinematic motions were compared. Cartilage pins were punched from the femoral condyles of porcine knee joints, and Co-Cr alloy disks were machined from orthopedic-grade rods and polished to a surface roughness ($R_a$) of 0.002. Friction tests were conducted by using a pin-on-disk-type tribotester in phosphate buffered saline (PBS) under pressures of 0.5, 1, and 2 MPa. All tests were performed in the repeat pass rotational (ROT) and the linear reciprocal (RCP) sliding motions with the same sliding distance and speed of 50 mm/s. The coefficients of friction of the cartilage against the Co-Cr alloy increased with the sliding time in both kinematic motions for all contact pressures. The maximum coefficients of friction in RCP motion were 1.08, 2.82, and 1.96 times those in ROT motion for contact pressures of 0.5, 1, and 2 MPa, respectively. As the contact pressure increased, the coefficients of friction gradually increased in RCP motion, whereas they decrease and then increased in ROT motion. The interaction between the directional change of the shear stress and the orientation of collagen fiber in the superficial layer of the cartilage could affect the change in the frictional behaviors of the cartilage. A large difference in the coefficients of friction between the two kinematic motions could be interpreted as differences in the directional change of shear stress at the contact surface.

Fire Retardant Treatment to the Plywood with Di-ammonium Phosphate [(NH4)2HPO4] (II) - Effect of Platen Temperature on Bending Strength of Treated Plywoods - (제2인산(第二燐酸) 암모늄에 의(依)한 합판(合板)의 내화처리(耐火處理)(II) - 열판온도(熱板溫度)가 처리합판(處理合板)의 곡강도(曲强度)에 미치는 영향(影響) -)

  • Chung, Woo-Yang;Lee, Phil-Woo
    • Journal of the Korean Wood Science and Technology
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    • v.12 no.2
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    • pp.3-9
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    • 1984
  • This study was carried out to examine the practicality of DAP[$(NH_4)_2HPO_4$] as fire retardant for plywood by static bending test the redried plywoods which had been soaked in 20% $(NH_4)_2HPO_4$ solution. Being hot/cold soaked in the solution for 3/3, 6/3, 9/3 and 12/3 hours and redried by cyclic press-drying method at the platen temp. of 130, 145, 100 and $175^{\circ}C$, the treated plywoods were tested to offer the mechanical data, that is, $S_{pl}$(stress at proportional limit), MOE(modulus of elasticity), MOR(modulus of rupture) and $W_{pl}$(work per unit volume to proportional limit ) in flexure. The results obtained were summarized as follows. 1. $S_{pl}$ of fire retardant treated plywoods ("FRP" would be used hereinafter) decreased as the platen temperature increased, but it was superior to that of non-treated plywoods(Control) at $160^{\circ}C$ or higher. 2. MOE of FRP decreased roughly with the increase of temperature, hut this tendency was not constant. And the value of FRP was higher than that of Control even at $175^{\circ}C$. 3. MOR of FRP showed same temperature-dependent tendency as MOE, but it was influenced more sensitively at the higher temperature. 4. $W_{pl}$ of FRP also decreased gradually with the increase of platen temperature and the value in DAP 9/3 treatment was Jess than 70% of control plywoods. 5. In view of redrying time and mechanical properties, the most reasonable platen temperature for DAP treated FRP was $160^{\circ}C$ in this study.

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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.