• Title/Summary/Keyword: Redox conditions

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Aspartate-glutamate carrier 2 (citrin): a role in glucose and amino acid metabolism in the liver

  • Milan Holecek
    • BMB Reports
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    • v.56 no.7
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    • pp.385-391
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    • 2023
  • Aspartate-glutamate carrier 2 (AGC2, citrin) is a mitochondrial carrier expressed in the liver that transports aspartate from mitochondria into the cytosol in exchange for glutamate. The AGC2 is the main component of the malate-aspartate shuttle (MAS) that ensures indirect transport of NADH produced in the cytosol during glycolysis, lactate oxidation to pyruvate, and ethanol oxidation to acetaldehyde into mitochondria. Through MAS, AGC2 is necessary to maintain intracellular redox balance, mitochondrial respiration, and ATP synthesis. Through elevated cytosolic Ca2+ level, the AGC2 is stimulated by catecholamines and glucagon during starvation, exercise, and muscle wasting disorders. In these conditions, AGC2 increases aspartate input to the urea cycle, where aspartate is a source of one of two nitrogen atoms in the urea molecule (the other is ammonia), and a substrate for the synthesis of fumarate that is gradually converted to oxaloacetate, the starting substrate for gluconeogenesis. Furthermore, aspartate is a substrate for the synthesis of asparagine, nucleotides, and proteins. It is concluded that AGC2 plays a fundamental role in the compartmentalization of aspartate and glutamate metabolism and linkage of the reactions of MAS, glycolysis, gluconeogenesis, amino acid catabolism, urea cycle, protein synthesis, and cell proliferation. Targeting of AGC genes may represent a new therapeutic strategy to fight cancer.

Backbone Cyclization of Flavin Mononucleotide-Based Fluorescent Protein Increases Fluorescence and Stability

  • Tingting Lin;Yuanyuan Ge;Qing Gao;Di Zhang;Xiaofeng Chen;Yafang Hu;Jun Fan
    • Journal of Microbiology and Biotechnology
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    • v.33 no.12
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    • pp.1681-1691
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    • 2023
  • Flavin mononucleotide-binding proteins or domains emit cyan-green fluorescence under aerobic and anaerobic conditions, but relatively low fluorescence and less thermostability limit their application as reporters. In this work, we incorporated the codon-optimized fluorescent protein from Chlamydomonas reinhardtii with two different linkers independently into the redox-responsive split intein construct, overexpressed the precursors in hyperoxic Escherichia coli SHuffle T7 strain, and cyclized the target proteins in vitro in the presence of the reducing agent. Compared with the purified linear protein, the cyclic protein with the short linker displayed enhanced fluorescence. In contrast, cyclized protein with incorporation of the long linker including the myc-tag and human rhinovirus 3C protease cleavable sequence emitted slightly increased fluorescence compared with the protein linearized with the protease cleavage. The cyclic protein with the short linker also exhibited increased thermal stability and exopeptidase resistance. Moreover, induction of the target proteins in an oxygen-deficient culture rendered fluorescent E. coli BL21 (DE3) cells brighter than those overexpressing the linear construct. Thus, the cyclic reporter can hopefully be used in certain thermophilic anaerobes.

Ecological Assessment of Plant Succession and Water Quality in Abandoned Rice Fields

  • Byun, Chae-Ho;Kwon, Gi-Jin;Lee, Do-Won;Wojdak, Jeremy M.;Kim, Jae-Geun
    • Journal of Ecology and Environment
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    • v.31 no.3
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    • pp.213-223
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    • 2008
  • The increasing area of abandoned rice fields could provide new opportunities for wetland restoration in Asia. However, it is unknown how quickly or completely abandoned rice fields will recover from agricultural disturbances. We assessed water quality and plant community succession in abandoned rice fields with different hydrology in a mountain valley to understand the effects of hydrological regime on recovery. Water level, soil redox potential, water quality, plant composition, and primary production were measured. The sites, coded as D6, N13, and N16, had been recovering for 6, 13, and 16 years by 2006. N13 and N16 have been recovering naturally whereas D6 has been drained with a nearby dike and was tilled in 2001. The typical hydroperiods of D6, N13, and N16 were no surface water, permanently flooded, and seasonally flooded, respectively. The major change in vegetation structure of both D6 and N13 was the replacement of herbaceous species by woody species. Drawdown accelerated this change because Salix koreensis grew better in damp conditions than in flooded conditions. Phragmites japonica reduced plot-level plant species richness. The removal efficiency of $NH_4-N$, $NO_3-N$, and $PO_4-P$ from water varied seasonally, ranging between -78.8 to 44.3%, 0 to 97.5%, and -26.0 to 44.4%, respectively. In summary, abandoned rice fields quickly became suitable habitat for native wetland plant species and improved regional water quality. Variation among our sites indicates that it is likely possible to manage abandoned rice fields, mostly through controlling hydrology, to achieve site-specific restoration goals.

Characterization of Nitrate Contamination and Hydrogeochemistry of Groundwater in an Agricultural Area of Northeastern Hongseong (홍성 북동부 농촌 지역 지하수의 질산성 질소 오염과 수리지구화학적 특성)

  • Ki, Min-Gyu;Koh, Dong-Chan;Yoon, Heesung;Kim, Hyun-Su
    • Journal of Soil and Groundwater Environment
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    • v.18 no.3
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    • pp.33-51
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    • 2013
  • Spatial and temporal characteristics of nitrate contamination and hydrogeochemical parameters were investigated for springs and surficial and bedrock groundwaters in northeastern part of Hongseong. Two field investigations were conducted at dry and wet seasons in 2011 for 120 sites including measurement of field parameters with chemical analyses of major dissolved constituents. Nitrate concentrations were at background levels in springs while 45% of bedrock groundwater and 49% of surficial groundwater exceeded the drinking water standard of nitrate (10 mg/L as $NO_3$-N). The difference in nitrate concentrations between surficial and bedrock groundwater was statistically insignificant. Cumulative frequency distribution of nitrate concentrations revealed two inflection points of 2 and 16 mg/L as $NO_3$-N. Correlation analysis of hydrogeochemical parameters showed that nitrate had higher correlations with Sr, Mg, Cl, Na, and Ca, in surficial groundwater in both dry and wet season. In contrast, nitrate had much weaker correlations with other hydrogeochemical parameters in bedrock groundwater compared to surficial groundwater and had significant correlations only in wet season. Temporally, nitrate and chloride concentrations decreased and dissolved oxygen (DO) increased from dry season to wet season, which indicates that increased recharge during the wet season affected groundwater quality. Aerobic conditions were predominant for both surficial and bedrock groundwater indicating low natural attenuation potential of nitrate in the aquifers of the study area.

Effects of chronic caloric restriction on kidney and heart redox status and antioxidant enzyme activities in Wistar rats

  • Dutra, Marcio Ferreira;Bristot, Ivi Juliana;Batassini, Cristiane;Cunha, Nubia Broetto;Vizuete, Adriana Fernanda Kuckartz;Souza, Daniela Fraga De;Moreira, Jose Claudio Fonseca;Goncalves, Carlos-Alberto
    • BMB Reports
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    • v.45 no.11
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    • pp.671-676
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    • 2012
  • Caloric restriction (CR) has been associated with health benefits and these effects have been attributed, in part, to modulation of oxidative status by CR; however, data are still controversial. Here, we investigate the effects of seventeen weeks of chronic CR on parameters of oxidative damage/modification of proteins and on antioxidant enzyme activities in cardiac and kidney tissues. Our results demonstrate that CR induced an increase in protein carbonylation in the heart without changing the content of sulfhydryl groups or the activities of superoxide dismutase and catalase (CAT). Moreover, CR caused an increase in CAT activity in kidney, without changing other parameters. Protein carbonylation has been associated with oxidative damage and functional impairment; however, we cannot exclude the possibility that, under our conditions, this alteration indicates a different functional meaning in the heart tissue. In addition, we reinforce the idea that CR can increase CAT activity in the kidney. Moreover, CR caused an increase in CAT activity in kidney, without changing other parameters. Protein carbonylation has been associated with oxidative damage and functional impairment; however, we cannot exclude the possibility that, under our conditions, this alteration indicates a different functional meaning in the heart tissue. In addition, we reinforce the idea that CR can increase CAT activity in the kidney.

pH Effect on the Structure of Reduced NifU-like Protein from Helicobacter pylori

  • Lee, Ki-Young;Kim, Ji-Hun;Bae, Ye-Ji;Lee, Bong-Jin
    • Journal of the Korean Magnetic Resonance Society
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    • v.19 no.3
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    • pp.106-111
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    • 2015
  • Helicobacter pylori (H. pylori) survives in acidic and fluctuating pH conditions of the stomach. The pH effect on H. pylori proteins is important for the advanced understanding of its evolution and viability, although this bacterium has the molecular machinery that neutralizes the acidic condition. HP1492 is known as a conserved NifU-like protein from H. pylori. NifU is a nitrogen fixation protein that mediates the transfer of iron-sulfur (Fe-S) cluster to iron-sulfur proteins like ferredoxin. Commonly, the monomeric reduced state of NifU can be converted to the dimeric oxidized state by intermolecular disulfide bond formation. Because it remains unclear that HP1492 actually behaves as known NifU protein, we first found that this protein can adopt both oxidized and reduced forms using size exclusion chromatography. Circular dichroism experiment showed that HP1492 is relatively well-structured at pH 6.5, compared to other pH conditions. On the basis of the backbone resonance assignment of HP1492, we further characterized the residues that are sensitive to pH using NMR spectroscopy. These residues showing large chemical shift changes could be mapped onto the secondary structure of the protein. Our results could provide the foundation for structural and biophysical studies on a wide spectrum of NifU proteins.

Analysis of the Growth and Metabolites of a Pyruvate Dehydrogenase Complex-Deficient Klebsiella pneumoniae Mutant in a Glycerol-Based Medium

  • Xu, Danfeng;Jia, Zongxiao;Zhang, Lijuan;Fu, Shuilin;Gong, Heng
    • Journal of Microbiology and Biotechnology
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    • v.30 no.5
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    • pp.753-761
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    • 2020
  • To determine the role of pyruvate dehydrogenase complex (PDHC) in Klebsiella pneumoniae, the growth and metabolism of PDHC-deficient mutant in glycerol-based medium were analyzed and compared with those of other strains. Under aerobic conditions, the PDHC activity was fourfold higher than that of pyruvate formate lyase (PFL), and blocking of PDHC caused severe growth defect and pyruvate accumulation, indicating that the carbon flux through pyruvate to acetyl coenzyme A mainly depended on PDHC. Under anaerobic conditions, although the PDHC activity was only 50% of that of PFL, blocking of PDHC resulted in more growth defect than blocking of PFL. Subsequently, combined with the requirement of CO2 and intracellular redox status, it was presumed that the critical role of PDHC was to provide NADH for the anaerobic growth of K. pneumoniae. This presumption was confirmed in the PDHC-deficient mutant by further blocking one of the formate dehydrogenases, FdnGHI. Besides, based on our data, it can also be suggested that an improvement in the carbon flux in the PFL-deficient mutant could be an effective strategy to construct high-yielding 1,3-propanediol-producing K. pneumoniae strain.

Construction of Microbial Fuel Cells Using Thermophilic Microorganisms, Bacillus licheniformis and Bacillus thermoglucosidasius

  • Choi, Young-Jin;Jung, Eun-Kyoung;Park, Hyun-Joo;Paik, Seung R.;Jung, Seun-Ho;Kim, Sung-Hyun
    • Bulletin of the Korean Chemical Society
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    • v.25 no.6
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    • pp.813-818
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    • 2004
  • A systematic study of microbial fuel cells comprised of thermophilic Bacillus licheniformis and Bacillus thermoglucosidasius has been carried out under various operating conditions. Substantial amount of electricity was generated when a redox mediator was used. Being affected by operation temperature, the maximum efficiency was obtained at 50$^{\circ}C$ with an open circuit voltage of ca. 0.7 V. While a small change around the optimum temperature did not make much effect on the cell performance, the rapid decrease in performance was observed above 70$^{\circ}C$. It was noticeable that fuel cell efficiency and discharge pattern strongly depended on the kind of carbon sources used in the initial culture medium. In the case of B. thermoglucosidasius, glucose alone was utilized constitutively as a substrate in the microbial fuel cell irrespective of used carbons sources. When B. licheniformis was cultivated with lactose as a carbon source, best charging characteristics were recorded. Trehalose, in particular, showed 41.2% coulombic efficiency when B. thermoglucosidasius was cultured in a starch-containing medium. Relatively good repetitive operation was possible with B. thermoglucosidasius cells up to 12 cycles using glucose as a carbon source, when they were cultured with lactose as an initial carbon source. This study demonstrates that highly efficient thermophilic microbial fuel cells can be constructed by a pertinent modulation of the operating conditions and by carefully selecting carbon sources used in the initial culture medium.

COMPARISON BETWEEN EXPERIMENTALLY MEASURED AND THERMODYNAMICALLY CALCULATED SOLUBILITIES OF UO2 AND THO2 IN KURT GROUND WATER

  • Kim, Seung-Soo;Baik, Min-Hoon;Kang, Kwang-Cheol;Choi, Jong-Won
    • Nuclear Engineering and Technology
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    • v.41 no.6
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    • pp.867-874
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    • 2009
  • Solubility of a radionuclide is important for defining the release source term of a radioactive waste in the safety and performance assessments of a radioactive waste repository. When the pH and redox potential of the KURT groundwater were changed by an electrical method, the concentrations of uranium and thorium released from $UO_2$(cr) and $ThO_2$(cr) at alkali pH(8.1 ${\sim}$ 11.4) and reducing potential (Eh < -0.2 V) conditions were less than $10^{-7}mole/L$. Unexpectedly, the concentration of tetravalent thorium is slightly higher than that of uranium at pH = 8.1 and Eh= -0.2 V conditions, and this difference may be due to the formation of hydroxide-carbonate complex ions. When $UO_2$(s) and $UO_2$(am, hyd.), and $ThO_2$(s) and $Th(OH)_4(am)$ were assumed as solubility limiting solid phases, the concentrations of uranium and thorium in the KURT groundwater calculated by the PHREEQC code were comparable to the experimental results. The dominating aqueous species of uranium and thorium were presumed as $UO_2(CO_3)_3^{4-}$ and $Th(OH)_3CO_3^-$ at pH = 8.1 ${\sim}$ 9.8, and $UO_2(OH)_3^-$ and $Th(OH)_4(aq)$ at pH = 11.4.

Regulation of Photosynthesis Genes (puf, puc, puhA, bchC, bchE, bchF, and bchI) in Rhodobacter sphaeroides (Rhodobacter sphaeroides에서의 광합성유전자(puf, puc, puhA, bchC, bchE, bchF와 bchI)의 발현조절)

  • Ko, In-Jeong;Kim, Yong-Jin;Lee, Jin-Mok;Shin, Sun-Joo;Oh, Jeong-Il
    • Journal of Life Science
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    • v.16 no.4
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    • pp.632-639
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    • 2006
  • Here we examined the expression patterns and regulation of seven photosynthesis (PS) genes (puf, puc, puhA, bchC, bchE, bchF, and bchI) in the anoxygenic photosynthetic bacterium, Rhodobacter sphaeroides, based on lacZ reporter gene assay. Expression of the tested PS genes, except puhA and bchI, were strongly induced in R. sphaeroides grown under anaerobic conditions relative to that under aerobic conditions. The puhA and bchI genes appear to form the operons together with bchFNBHLM-RSP0290 and crtA, respectively. Expression of the puf, puc, and bchCXYZ operons in R. sphaeroides grown photosynthetically was proportional to the incident light intensity, whereas that of bchFNBHLM(RSP0290-puhA) was inversely related to light intensity. Expression of bchEJG was lowest under medium-light photosynthetic conditions $(10\;W/m^2)$ and highest under high light conditions $(100\;W/m^2)$. The regulation of PS genes by the three major regulatory systems involved in oxygen- and light-sensing in R. sphaeroides is as following: puf and bchC are regulated by both the PpsR repressor and the PrrBA two-component system. The puc operon is under control of PpsR, FnrL, and PrrBA system. Expression of bchE is controlled by FnrL and PrrBA two-component system, whereas bchF is regulated exclusively by PpsR. It was demonstrated that the PpsR repressor is responsible for high-light repression of bchF and that FnrL might be involved in perceiving the cellular redox state in addition to sensing $O_2$ itself.