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1.
Both the changes in the activities of nitrogenase, glutamine synthetase and glutamate dehydrogenase and in the extracellular and intracellular NH4 + concentrations were investigated during the transition from an NH4 + free medium to one containing NH4 + ions for a continuous culture of Azotobacter vinelandii. If added in amounts causing 80–100% repression of nitrogenase, ammonium acetate, lactate and phosphate are absorbed completely, whereas chloride, sulfate and citrate are only taken up to about 80%. After about 1–2 hrs the NH4 + remaining in the medium is absorbed too, indicating the induction or activation of a new NH4 + transport system. One of the new permeases allows the uptake of citrate in the presence of sucrose. Addition of inorganic NH4 + salts leads to acidification of the culture. Anaerobiosis suppresses NH4 + transport. A rise in the extracellular NH4 + level leads to a reversible rise in the glutamine synthetase activity, which is not prevented by chloramphenicol, and to a reversible decrease in nitrogenase activity. During these measurements glutamate dehydrogenase activity remains close to zero. The intracellular NH4 + level of about 0.6 mM does not change when extracellular NH4 + is taken up and repression of nitrogenase starts.  相似文献   

2.
Both nitrite reductase and nitrate reductase were induced by nitrite, but there were differences in the time course of induction and in the response to different NO2 - concentrations between these enzymes. NH4 + depressed the induction of nitrite reductase. NADH2 dependent glutamate dehydrogenase activity was enhanced by those NO2-concentrations in the medium at which unmetabolized NO2 - occurred in the roots. NADPH2 and NAD+ dependent GDh activities were not affected. In vivo modification and (or) in vivo activation were probably responsible for the increase in NADH2 dependent GDH activity.  相似文献   

3.
Protoveratrine-(5 M) stimulated aerobic glycolysis of incubated rat brain cortex slices that accompanies the enhanced neuronal influx of Na+ is blocked by tetrodotoxin (3 M) and the local anesthetics, cocaine (0.1 mM) and lidocaine (0.5 mM). On the other hand, high [K+]-stimulated aerobic glycolysis that accompanies the acetylcholine-sensitive enhanced glial uptakes of Na+ and water is unaffected by acetylcholine (2 mM). Experiments done under a variety of metabolic conditions show that there exists a better correlation between diminished ATP content of the tissue and enhanced aerobic glycolysis than between tissue ATP and the ATP-dependent synthesis of glutamine. Whereas malonate (2 mM) and amino oxyacetate (5 mM) suppress ATP content and O2 uptake, stimulate lactate formation, but have little effect on glutamine levels, fluoroacetate (3 mM) suppresses glutamine synthesis in glia, presumably by suppressing the operation of the citric acid cycle, with little effect on ATP content, O2 uptake, and lactate formation. Exogenous citrate (5 mM), which may be transported and metabolized in glia but not in neurons, inhibits lactate formation by cell free acetone-dried powder extracts of brain cortex but not by brain cortex slices. These results suggest that the neuron is the major site of stimulated aerobic glycolysis in the brain, and that under our experimental conditions glycolysis in glia is under lesser stringent metabolic control than that in the neuron. Stimulation of aerobic glycolysis by protoveratrine occurs due to diminution of the energy charge of the neuron as a result of stimulation of the sodium pump following tetrodotoxin-sensitive influx of Na+; stimulation by high [K+, NH4 +, or Ca2+ deprivation occurs partly by direct stimulation of key enzymes of glycolysis and partly by a fall in the tissue ATP concentration.  相似文献   

4.
We studied the influence of inorganic nitrogen sources (NO3 ? or NH4 +) and potassium deficiency on expression and activity of plasma membrane (PM) H+-ATPase in sorghum roots. After 15 d of cultivation at 0.2 mM K+, the plants were transferred to solutions lacking K+ for 2 d. Then, K+ depletion assays were performed in the presence or absence of vanadate. Further, PMs from K+-starved roots were extracted and used for the kinetic characterization of ATP hydrolytic activity and the immunodetection of PM H+-ATPase. Two major genes coding PM H+-ATPase (SBA1 and SBA2) were analyzed by real-time PCR. PM H+-ATPase exhibited a higher Vmax and Km in NH4 +-fed roots compared with NO3 ? -fed roots. The optimum pH of the enzyme was slightly lower in NO3 ? -fed roots than in NH4 +-fed roots. The vanadate sensitivity was similar. The expressions of SBA1 and SBA2 increased in roots grown under NH4 +. Concomitantly, an increased content of the enzyme in PM was observed. The initial rate of K+ uptake did not differ between plants grown with NO3 ? or NH4 +, but it was significantly reduced by vanadate in NH4 +-grown plants.  相似文献   

5.
Three halophilic mycelium-forming actinobacteria, strains H195T, H150 and H151, were isolated from a Saharan soil sample collected from Béni-isguen in the Mzab region (Ghardaïa, South of Algeria) and subjected to a polyphasic taxonomic characterisation. These strains were observed to show an aerial mycelium differentiated into coccoid spore chains and fragmented substrate mycelium. Comparative analysis of the 16S rRNA gene sequences revealed that the highest sequence similarities were to Saccharopolyspora qijiaojingensis YIM 91168T (92.02 % to H195T). Phylogenetic analyses showed that the strains H195T, H150 and H151 represent a distinct phylogenetic lineage. The cell-wall hydrolysate was found to contain meso-diaminopimelic acid, and the diagnostic whole-cell sugars were identified as arabinose and galactose. The major cellular fatty acids were identified as iso-C15:0, iso-C16:0, iso-C17:0 and anteiso-C17:0. The diagnostic phospholipid detected was phosphatidylcholine and MK-9 (H4) was found to be the predominant menaquinone. The genomic DNA G+C content of strain H195T was 68.2 mol%. On the basis of its phenotypic features and phylogenetic position, we propose that strain H195T represents a novel genus and species, Mzabimyces algeriensis gen. nov., sp. nov., within a new family, Mzabimycetaceae fam. nov. The type strain of M. algeriensis is strain H195T (=DSM 46680T = MTCC 12101T).  相似文献   

6.
The proton/hydroxide (H+/OH) permeability of phospholipid bilayer membranes at neutral pH is at least five orders of magnitude higher than the alkali or halide ion permeability, but the mechanism(s) of H+/OH transport are unknown. This review describes the characteristics of H+/OH permeability and conductance through several types of planar phospholipid bilayer membranes. At pH7, the H+/OH conductances (G H/OH) range from 2–6 nS cm–2, corresponding to net H+/OH permeabilities of (0.4–1.7)×10–5 cm sec–1. Inhibitors ofG H/OH include serum albumin, phloretin, glycerol, and low pH. Enhancers ofG H/OH include chlorodecane, fatty acids, gramicidin, and voltages >80 mV. Water permeability andG H/OH are not correlated. The characteristics ofG H/OH in fatty acid (weak acid) containing membranes are qualitatively similar to the controls in at least eight different respects. The characteristics ofG H/OH in gramicidin (water wire) containing membranes are qualitatively different from the controls in at least four different respects. Thus, the simplest explanation for the data is thatG H/OH in unmodified bilayers is due primarily to weakly acidic contaminants which act as proton carriers at physiological pH. However, at low pH or in the presence of inhibitors, a residualG H/OH remains which may be due to water wires, hydrated defects, or other mechanisms.  相似文献   

7.
Fructose reacts spontaneously with proteins in the brain to form advanced glycation end products (AGE) that may elicit neuroinflammation and cause brain pathology, including Alzheimer's disease. We investigated whether fructose is eliminated by oxidative metabolism in neocortex. Injection of [14C]fructose or its AGE‐prone metabolite [14C]glyceraldehyde into rat neocortex in vivo led to formation of 14C‐labeled alanine, glutamate, aspartate, GABA, and glutamine. In isolated neocortical nerve terminals, [14C]fructose‐labeled glutamate, GABA, and aspartate, indicating uptake of fructose into nerve terminals and oxidative fructose metabolism in these structures. This was supported by high expression of hexokinase 1, which channels fructose into glycolysis, and whose activity was similar with fructose or glucose as substrates. By contrast, the fructose‐specific ketohexokinase was weakly expressed. The fructose transporter Glut5 was expressed at only 4% of the level of neuronal glucose transporter Glut3, suggesting transport across plasma membranes of brain cells as the limiting factor in removal of extracellular fructose. The genes encoding aldose reductase and sorbitol dehydrogenase, enzymes of the polyol pathway that forms glucose from fructose, were expressed in rat neocortex. These results point to fructose being transported into neocortical cells, including nerve terminals, and that it is metabolized and thereby detoxified primarily through hexokinase activity.

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8.
Methane formation from H2/CO2 by methanogenic bacteria is dependent on Na+ ions. In this communication it is shown with Methanobacterium thermoautotrophicum that a Na+/H+ antiporter plays a role in methane formation from H2 and CO2 and in the regulation of the ΔpH. This is based on the following findings:
  1. Li+ ions, an alternative substrate of Na+/H+ antiporters, could replace Na+ in stimulating methanogenesis from H2 and CO2.
  2. Harmaline, amiloride, and NH 4 + , which are inhibitors of Na+/H+ antiporters, inhibited methanogenesis; inhibition was competitive to Na+ or Li+.
  3. Addition of Na+ or Li+ rather than of other cations to cell suspensions resulted in an acidification of the suspension medium. The rate and extent of acidification was affected by those inhibitors, which inhibited methanogenesis competitively to Na+ or Li.
  4. During methane formation from H2 and CO2 the generation of a ΔpH (inside alkaline) was dependent on the presence of Na+ or Li+. However, methanogenesis was also dependent on Na+ or Li+ under conditions where ΔpH was zero.
  5. ATP synthesis driven by an electrogenic potassium efflux was significantly enhanced in the presence of Na+ or Li+. Na+ or Li+ were shown to prevent acidification of the cytoplasm under these conditions.
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9.
The growth yields of three strains of Rhizobium japonicum (CB 1809, CC 723, CC 705) in culture solutions containing L-glutamate were about twice those grown with ammonium. The activities of glutamine synthetase (GS; EC 6.3.1.2) and glutamate dehydrogenase (GDH; EC 1.4.1.4) were dependent on the nitrogen source in the medium and also varied with growth. Both NADPH-and NADH-dependent glutamate synthase (GOGAT; EC 1.4.1.13) and NADPH-dependent GDH were found in strains grown with either glutamate or ammonium but NADH-linked GDH was only detected in glutamate-grown cells. Glutamine synthetase was adenylylated in cells grown with NH 4 + (90%) and to lesser extent in those grown with L-glutamate (50%). In root nodules produced by the three strains in Glycine max (L.) Merr., the bulk of GS was located in the nodule cytosol (60–85%). The enzyme was adenylylated in bacteroids (43–75%) and in the nodule tissues (52–68%). The enzyme in cell-free extracts of Rh. japonicum (CC 705) grown in culture solutions containing glutamate and in bacteroids (CC 705) was deadenylylated by snake-venom phosphodiesterase. L-methionine-DL-sulfoximine restricted the incoporation of 15N-labelled (NH4)2SO4 into cells of strains CB 1809 and CC 705, as well as in bacteroids of strain CC 705. It is noteworthy that appreciable activities for GDH were found in the free-living rhizobia grown on glutamate. Thus the presence of an enzyme does not necessarily imply that a particular pathway is operative in assimilating ammonium into cell nitrogen. Based on 15N studies, the GS-GOGAT pathway of rhizobia (strains CB 1809 and CC 705) is important when grown in culture solutions as well as in bacteroids from root nodules of G. max.  相似文献   

10.
We investigated effects of Ebselen, diphenyl diselenide (PhSe)2 and diphenyl ditelluride (PhTe)2 on [3H]glutamate uptake and release by brain synaptosomes. Ebselen after acute exposure inhibited K+-stimulated [3H]glutamate release by brain synaptosomes. (PhSe)2 and (PhTe)2 did not change [3H]glutamate release by brain synaptosomes. Ebselen, (PhSe)2 and (PhTe)2 had no significantly effects on [3H]glutamate uptake after acute exposure. In vitro, Ebselen (100 M) inhibited [3H]glutamate release and uptake. (PhSe)2 had no significant effect, while (PhTe)2 (100 M) inhibited [3H]glutamate uptake by brain synaptosomes. In vitro, (PhSe)2, (PhTe)2 and Ebselen caused a significant inhibition of [3H]glutamate uptake by brain synaptic vesicles in vitro. The results demonstrated that organochalcogenides have a rather complex effect on glutamate homeostasis depending on the compound and the schedule of exposition. We propose that the neuroprotective action of Ebselen can be related, in addition to its glutathione peroxidase-like and antilipoperoxidative activity, to a direct interaction with the glutamatergic system by reducing Kï-evoked glutamate release.  相似文献   

11.
The effect of ion fluxes across the inner membrane on calcium-independent uncoupling activity of palmitic acid was investigated in experiments on rat liver mitochondria energized by the oxidation of succinate. The following compounds were used as the inductors of ion fluxes: the K+/H+ antiporter nigericin causing transformation of ΔpH into electrical potential difference (Δψ) across the inner membrane; tetraphenylphosphonium (TPP+) that freely crosses phospholipid membranes; protonophore carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) that induces a flow of H+ from the intermembrane space into the matrix and reduces Δψ and ΔpH. It was found that nigericin at a concentration of 20 nM, which causes an increase in maximal Δψ, partially inhibits the ability of palmitic acid to reduce Δψ and stimulates mitochondrial respiration. A specific inhibitor of the ATP/ADP antiporter (carboxyatractylate) and a substrate of the aspartate/glutamate antiporter (glutamate) increase Δψ and partially inhibit mitochondrial respiration in the presence of palmitic acid. Under these conditions, 10 μM cyclosporin A also inhibits respiration but has no effect on Δψ. The specific uncoupling activity of palmitic acid (V U) and its specific components that characterize participation of the ATP/ADP antiporter (V Catr), aspartate/glutamate antiporter (V Glu), and cyclosporin-A-sensitive system (V CsA) in the palmitic acid-induced uncoupling were estimated. It was shown that nigericin substantially reduces V U, V Catr and V Glu but increases V CsA. TPP+ at a concentration of 20 μM increases V U and V Glu, does not affect V Catr and reduces V CsA. FCCP at concentrations of 20 and 40 nM reduces Δψ by not more than 17% but does not affect V U, V Catr, V Glu and V CsA. The results suggest that the calcium-independent uncoupling effect of palmitic acid in liver mitochondria is caused by the return of protons to the matrix with participation of ADP/ATP and aspartate/glutamate antiporters and owing to activation of cyclosporin A-sensitive electron transport along the respiratory chain without affecting Δψ. The induced ion fluxes across the inner mitochondrial membrane can be considered as a factor of the calcium-independent regulation of uncoupling activity of palmitic acid in liver mitochondria with participation of the ADP/ATP and aspartate/glutamate antiporters and of the cyclosporin A-sensitive electron transport system.  相似文献   

12.
Binding ofl-[3H]glutamate to membranes from whole chick retina and from subcellular fractions enriched with photoreceptor terminals (P1), or terminals from the inner plexiform layer (P2) was studied. Na+-dependent and Na+-independent binding to these membranes was demonstrated. Na+-independent binding was stereospecific. Kinetic analysis of the binding process indicated a single high-affinity system (K B=0.55 M) with a capacity of approximately 20 pmoles/mg protein in all the membrane fractions. [3H]Glutamate binding to P1 and P2 fractions was effectively displaced by several structural analogues of glutamate. Glutamate diethyl-ester appreciably displaced binding, whereas kainic acid did not displace bound glutamate. Data indicate the binding of [3H]glutamate to physiologically relevant receptors in the chick retina.  相似文献   

13.
A Gram-negative bacterial strain, designated WB1T, was isolated from a domestic refrigerator in Guangzhou, PR China. Cells of strain WB1T were oxidase-negative, catalase-positive, strictly aerobic, non-spore-forming and non-motile coccobacilli with peritrichous fimbriae-like structures. The strain was able to grow at 10–40 °C with optimum growth at 28–30 °C, pH 6.0–8.0 (optimum, pH 7.0) and 0–6 % NaCl (w/v, optimum, 0.5 %). Phylogenetic analyses based on 16S rRNA gene and rpoB gene sequences revealed that strain WB1T belonged to the genus Acinetobacter and was most closely related to A. indicus DSM 25388T (97.2 % 16S rRNA gene sequence similarity) and A. radioresistens NBRC 102413T (96.8 %). The DNA G + C content of strain WB1T was 46.74 ± 0.04 mol % and the major fatty acids comprised summed feature 3 (C16:1 ω7c and/or C16:1 ω6c), C18:1 ω9c, C16:0 and C12:0. The predominant respiratory quinone was identified as Q-9 and the polar lipids as diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine and an unidentified phospholipid. Phenotypic, phylogenetic and chemotaxonomic data, including low DNA–DNA relatedness with closely related type strains, supported that strain WB1T represents a distinct novel species in the genus Acinetobacter, for which the name Acinetobacter refrigeratorensis sp. nov. was proposed. The type strain is WB1T (=GIMCC 1.663T = CCTCC AB 2014197T = KCTC 42011T).  相似文献   

14.
Our previous work revealed proanthocyanidins (PAs) could pose significant enhancement on the activity of H+-ATPase and fermentation efficiency after a transient initial inhibition (Li et al in Am J Enol Vitic 62(4):512–518, 2011). The aim of the present work was to understand the possible mechanism for this regulation. At Day 0.5 the gene expression level of PMA1 in AWRI R2 strain supplemented with 1.0 mg/mL PAs was decreased by around 54 % with a 50 % and a 56.5 % increase in the concentration of intracellular ATP and NADH/NAD+ ratio, respectively, compared to that of control. After the transient adaptation, the gene expression levels of PMA1 and HXT7 in PAs-treated cells were enhanced significantly accompanied by the decrease of ATP contents and NADH/NAD+ ratio, which resulted in the high level of the activities of rate-limiting enzymes. PAs could pose significant effects on the fermentation via glucose transport, the energy and redox homeostasis as well as the activities of rate-limiting enzymes in glycolysis.  相似文献   

15.
Hydroxylamine added to the nutrient medium in sublethal concentrations (0.2 to 1.0 mN) enhanced NADH2 dependent glutamate dehydrogenase activity in isolated pea roots. The increase in activity depended on proteosynthesis and was lower in the presence of NO3 ? and NH4 + ions. The induction of nitrate reductase and of nitrite reductase was partly inhibited by sublethal hydroxylamine concentrations.  相似文献   

16.
Summary The energetics, enzyme activities and end-product synthesis of Zymomonas mobilis 113 in continuous culture were studied after the shift from an anaerobic to an aerobic environment. Aeration diminished ethanol yield and lactic acid concentration, but increased glucose consumption rate and production of acetic acid. After the shift to aerobic conditions reduced nicotinamide adenine dinucleotide (phosphate) [NAD(P)H]-oxidase activity was stimulated. Washed cell suspensions consumed oxygen with glucose, lactate and ethanol as substrates. The aerobic Z. mobilis 113 regulated their intracellular redox balance by production and reoxidation of the end products, coupled with the formation of NAD(P)H. An increase in transmembrane pH gradient (pH) and a decrease in intracellular ATP concentration were observed after the shift to aerobic conditions. At low medium redox potential (Eh) values the H+ balance was regulated in an energy-independent way via end-product excretion. Under aerobic conditions this was supplemented by ATP-dependent H+ excretion by the membrane H+-ATPase.Abbreviations D dilution rate (h-1) - S 0 initial glucose concentration (g/l) - Y x/s growth yield (g/mol) - Y p/s product yield (g/g) - q s specific rate of substrate utilization (g/g per hour) - q p specific rate of ethanol formation (g/g per hour) - qo 2 specific rate of CO2 production (mmol/g per hour) - specific growth rate (h-1) - X dry biomass concentration (g/l) - Eh redox potential of culture medium (mV) - pH transmembrane pH gradient (pH units) - pHin intracellular pH - SASE sum of activities of specific enmymes of Entner-Doudoroff pathway  相似文献   

17.
Kinetics of the separation of L-glutamic acid (GLU) by ion exchange has been studied with strongly acidic H+-type cation exchange resin Amberlite IR-122. Since glutamic acid is a trivalent ampholyte and dissociates according to three equilibrium reactions, separation of G+ ions by a cation exchange process is accompanied by the dominant reversible reaction, i.e. G++H+ ? G0. Accompanying reversible reaction has an effect on the ion exchange rate, and decreases the performance of the process comparing with the ideal case that the exchanging ions retain their identity. The analysis was performed first with the ion exchange column, DIC (L/D=0.52); and then with the ion exchange column, IC (L/D=10.9). The data were collected with model glutamic acid solutions for both DIC and IC columns/reactors. IC experimental results were then compared with that of DIC and the effect of scale up on ion exchange process was investigated. The experimental results have provided an adequate basis for the design calculations, and the design parameters were determined. Rate coefficients for the liquid phase mass transfer controlled cation exchange process were calculated and interrelated with a plot of j Mfactor versus Reynolds number.  相似文献   

18.
A Gram-stain-negative, aerobic, non-motile and rod-shaped or ovoid bacterial strain, GJSW-22T, which was isolated from seawater at Geoje island in South Korea, was characterized taxonomically. Strain GJSW-22T was observed to grow optimally at 30 °C, at pH 7.0–8.0 and in the presence of 2.0 % (w/v) NaCl. In the neighbour-joining phylogenetic tree based on 16S rRNA gene sequences, strain GJSW-22T grouped with the type strains of Thalassobius species, forming a stable cluster with the type strain of Thalassobius aestuarii (bootstrap value of 83.2 %). Strain GJSW-22T exhibited the highest 16S rRNA gene sequence similarity value (98.0 %) to the type strain of T. aestuarii. It exhibited 16S rRNA gene sequence similarity values of 95.6–96.1 % to the type strains of the other Thalassobius species. Strain GJSW-22T was found to contain Q-10 as the predominant ubiquinone and C18:1 ω7c and 11-methyl C18:1 ω7c as the major fatty acids. The major polar lipids of strain GJSW-22T were identified as phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, one unidentified aminolipid and one unidentified lipid. The DNA G + C content of strain GJSW-22T is 60.3 mol % and its mean DNA–DNA relatedness value with the type strain of T. aestuarii was 23 %. Differential phenotypic properties, together with the phylogenetic and chemotaxonomic data, confirmed that strain GJSW-22T is distinct from other Thalassobius species. On the basis of the data presented, strain GJSW-22T is considered to represent a novel species of the genus Thalassobius, for which the name Thalassobius aquaeponti sp. nov. is proposed. The type strain is GJSW-22T (=KCTC 42115T = NBRC 110378T).  相似文献   

19.
20.
A Gram-stain negative, non-flagellated, non-gliding, aerobic and rod-shaped bacterium, designated GYSW-15T, was isolated from a seawater of Geoje island on the South Sea, South Korea, and subjected to a polyphasic taxonomic approach. Strain GYSW-15T was found to grow optimally at 30 °C, at pH 7.0–8.0 and in the presence of approximately 2.0 % (w/v) NaCl. Neighbour-joining phylogenetic tree based on 16S rRNA gene sequences showed that strain GYSW-15T belongs to the genus Polaribacter, clustering coherently with the type strain of Polaribacter porphyrae showing sequence similarity value of 97.0 %. Strain GYSW-15T exhibited 16S rRNA gene sequence similarity values of 94.3–96.4 % to the type strains of the other Polaribacter species. Strain GYSW-15T was found to contain MK-6 as the predominant menaquinone and iso-C15:0, iso-C15:1 G, iso-C16:0 3-OH and summed feature 3 (C16:1 ω7c and/or C16:1 ω6c) as the major fatty acids. The polar lipid profile of strain GYSW-15T containing phosphatidylethanolamine and an unidentified lipid as major components was similar to that of the type strain of P. porphyrae. The DNA G + C content of strain GYSW-15T was determined to be 31.2 mol% and its mean DNA–DNA relatedness with P. porphyrae LMG 26671T was 17 %. Differential phenotypic properties, together with its phylogenetic and genetic distinctiveness, revealed that strain GYSW-15T is separated from recognized Polaribacter species. On the basis of the data presented, strain GYSW-15T is considered to represent a novel species of the genus Polaribacter, for which the name Polaribacter marinivivus sp. nov. is proposed. The type strain is GYSW-15T ( = KCTC 42156T = CECT 8655T)  相似文献   

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