Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:1.7.1.2 (nitrate reductase)
 3,861 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The "24-HOUR BATH" is an apparatus which circulates the bath water, keeps it clean and warm, and makes it possible to take a bath at any time during the day or night. It consists of apparatus for cleaning (sponge or mesh filter and filter material), heating (ceramic heater), and sterilizing (UV lamp). Recently, three cases of skin disease due to M. avium infection in private homes, in which "24-HOUR BATH" water was suspected to be the source of infection, have been reported. We attempted to isolate M. avium complex from the water (32 specimens), sponge filter (29 specimens), and filter material (32 specimens) of the "24-HOUR BATH". One hundred-ml samples of bath water, and 50-ml samples of rinse from a sponge filter or filter material were centrifuged at 3000 rpm for 20 min. Sediment was suspended in distilled water and a smear was prepared, and then digested and decontaminated with 2% sodium hydroxide. The processed specimens were cultured on 2% Ogawa medium containing ofloxacin (1 microgram/ml) and ethambutol (2.5 micrograms/ml) for 8 weeks at 37 degrees C. Positive smears were 3 (9.4%), 25 (86.2%) and 25 (78.1%) specimens from the water, sponge and filter material, respectively. A few bacterial clumps were observed, especially in the sponge specimens. The number of positive culture was 5 (15.6%), 24 (82.8%) and 25 (78.1%) from the water, sponge and filter material, respectively. Among them the number of Runyon's Group III-positive cultures was 5 (100%), 22 (91.7%) and 20 (80%) in the water, sponge, and filter material specimens, respectively. In most cases, cultures were positive for both the sponge and filter material specimens. All of the Group III mycobacteria were smooth, grew at 28, 37, 42, and 45 degrees C, negative for niacin, nitrate reductase, semiquantitative catalase, urease and Tween80 hydrolysis, and positive for 68 degrees C catalase. All of the strains reacted with M. avium complex AccuProbe and M. avium AccuProbe, but none of the strains reacted with M. intracellulare AccuProbe. Therefore, all the Group III isolates were identified as M. avium by the culture, biochemical and genetical characteristics.
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PMID:[Isolation of Mycobacterium avium complex from the "24-hour bath"]. 1068 14

The presence of NaCl in the nutrient solution promoted nitrate uptake in parent Anabaena sp. PCC 7120, mutants SP7 (defective in nitrate reductase activity) and SP17 (partially defective in nitrate reductase activity), but not in the mutant SP9 (defective in nitrate transport and reduction). Nitrate reductase activity of the parent and mutant SP17 increased with increasing concentration of nitrate in saline medium, while mutants SP7 and SP9 did not respond to the altered salinity. Although Na+ was not required for nitrate reductase activity, its presence in the nutrient solution enhanced nitrate reduction. Complete removal of Na+ from the nutrient solution markedly reduced nitrogenase activity in all the strains, while raising the concentration of NaCl to 50 mmol l-1 or above, was equally toxic to nitrogenase activity. External NaCl at 200 mmol l-1 brought down the nitrogenase activity to the same residual level as observed without Na+.
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PMID:Response to NaCl of nitrate assimilation and nitrogenase activity of the cyanobacterium Anabaena sp. PCC 7120 and its mutants. 1069 73

The transcription factor NNR from Paracoccus denitrificans was expressed in a strain of Escherichia coli carrying a plasmid-borne fusion of the melR promoter to lacZ, with a consensus FNR-binding site 41.5 bp upstream of the transcription start site. This promoter was activated by NNR under anaerobic growth conditions in media containing nitrate, nitrite, or the NO(+) donor sodium nitroprusside. Activation by nitrate was abolished by a mutation in the molybdenum cofactor biosynthesis pathway, indicating a requirement for nitrate reductase activity. Activation by nitrate was modulated by the inclusion of reduced hemoglobin in culture media, because of the ability of hemoglobin to sequester nitric oxide and nitrite. The ability of nitrate and nitrite to activate NNR is likely due to the formation of NO (or related species) during nitrate and nitrite respiration. Amino acids potentially involved in NNR activity were replaced by site-directed mutagenesis, and the activities of NNR derivatives were tested in the E. coli reporter system. Substitutions at Cys-103 and Tyr-35 significantly reduced NNR activity but did not abolish the response to reactive nitrogen species. Substitutions at Phe-82 and Tyr-93 severely impaired NNR activity, but the altered proteins retained the ability to repress an FNR-repressible promoter, so these mutations have a "positive control" phenotype. It is suggested that Phe-82 and Tyr-93 identify an activating region of NNR that is involved in an interaction with RNA polymerase. Replacement of Ser-96 with alanine abolished NNR activity, and the protein was undetectable in cell extracts. In contrast, NNR in which Ser-96 was replaced with threonine retained full activity.
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PMID:Heterologous NNR-mediated nitric oxide signaling in Escherichia coli. 1105 88

Measurement of nitrite and nitrate, the stable oxidation products of nitric oxide (NO), provides a useful tool to study NO synthesis in vivo and in cell cultures. A simple and rapid fluorometric HPLC method was developed for determination of nitrite through its derivatization with 2,3-diaminonaphthalene (DAN). Nitrite, in standard solution, cell culture medium, or biological samples, readily reacted with DAN under acidic conditions to yield the highly fluorescent 2,3-naphthotriazole (NAT). For analysis of nitrate, it was converted to nitrite by nitrate reductase, followed by the derivatization of nitrite with DAN to form NAT. NAT was separated on a 5-microm reversed-phase C18 column (150X4.6 mm, I.D.) guarded by a 40-microm reversed-phase C18 column (50x4.6 mm, I.D.), and eluted with 15 mM sodium phosphate buffer (pH 7.5) containing 50% methanol (flow-rate, 1.3 ml/min). Fluorescence was monitored with excitation at 375 nm and emission at 415 nm. Mean retention time for NAT was 4.4 min. The fluorescence intensity of NAT was linear with nitrite or nitrate concentrations ranging from 12.5 to 2,000 nM in water, cell culture media, plasma and urine. The detection limit for nitrite and nitrate was 10 pmol/ml. Because NAT is well separated from DAN and other fluorescent components present in biological samples, our HPLC method offers the advantages of high sensitivity and specificity as well as easy automation for quantifying picomole levels of nitrite and nitrate in cell culture medium and biological samples.
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PMID:Rapid determination of nitrite by reversed-phase high-performance liquid chromatography with fluorescence detection. 1107 72

The napB gene of the pathogenic bacterium Haemophilus influenzae encodes a dihaem cytochrome c, the small subunit of a heterodimeric periplasmic nitrate reductase (Nap). Recombinant NapB was overproduced in Escherichia coli, purified to near-homogeneity and crystallized using the hanging-drop method. Thin quadrilateral plates were grown under various conditions but proved to be unsuitable for X-ray analysis. However, a single crystal was grown using 1.75 M ammonium sulfate in 0.1 M sodium acetate pH 5.5, from which a native data set could be collected to 1.8 A resolution using synchrotron radiation. Using the same conditions, further crystals were obtained by microseeding. The space group was determined to be P42(1)2, with unit-cell parameters a = 77.55, b = 77.55, c = 28.64 A and an unusually low solvent content of 16.5%, assuming there to be one molecule of NapB in the asymmetric unit. Analysis of the dissolved crystals indicated that partial proteolysis of the protein had occurred. Taking the molecular mass of the crystallized form ( approximately 8500 Da) into account, the solvent content was estimated to be 53%, with a V(M) value of 2.64 A(3) Da(-1).
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PMID:Crystallization and preliminary X-ray analysis of the recombinant dihaem cytochrome c (NapB) from Haemophilus influenzae. 1122 19

Salmonella cause economic losses to the swine industry due to disease and compromised food safety. Since the gut is a major reservoir for Salmonella, strategies are sought to reduce their concentration in pigs immediately before processing. Respiratory nitrate reductase activity possessed by Salmonella also catalyzes the intracellular reduction of chlorate (an analog of nitrate) to chlorite, which is lethal to the microbe. Since most gastrointestinal anaerobes lack respiratory nitrate reductase, we conducted a study to determine if chlorate may selectively kill Salmonella within the pig gut. Weaned pigs orally infected with 8 x 10(7) CFU of a novobiocin- and nalidixic acid-resistant strain of Salmonella Typhimurium were treated 8 and 16 h later via oral gavage (10 ml) with 0 or 100 mM sodium chlorate. Pigs were euthanized at 8-h intervals after receiving the last treatment. Samples collected by necropsy were cultured qualitatively and quantitatively for Salmonella and for most probable numbers of total culturable anaerobes. A significant (P < 0.05) chlorate treatment effect was observed on cecal concentrations of Salmonella, with the largest reductions occurring 16 h after receiving the last chlorate treatment. An observed treatment by time after treatment interaction suggests the chlorate effect was concentration dependent. Chlorate treatment may provide a means to reduce foodborne pathogens immediately before harvest.
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PMID:Effect of sodium chlorate on Salmonella typhimurium concentrations in the weaned pig gut. 1127 77

An integrated enzyme-functionalized field-effect transistor (ENFET) device for the sensing of nitrate ions is described. An aminosiloxane-functionalized gate interface is modified with N-methyl-N'-(carboxyalkyl)-4,4'-bipyridinium relay units. The complex formed between nitrate reductase and the bipyridinium units on the gate surface is crosslinked with glutaric dialdehyde to yield a stable relay-enzyme layer on the gate interface. In the presence of sodium dithionite as electron donor, the biocatalyzed reduction of nitrate to nitrite ion is stimulated. The ratio between the oxidized and reduced states of the bipyridinium sites regulates the gate potential, and is controlled by the concentration of NO3- ions in the system. The effect of the chain length tethering the N-methyl-N'-(carboxyalkyl)-4,4'-bipyridinium units to the gate surface on the biocatalyzed reduction of NO3- ions, and on the NO3- FET sensor performance is discussed. The devices that include the bipyridinium units tethered to the gate interface with methylene chain length, -(CH2)n, where n > or = 7, reveal a detection limit of 7 x 10(-5) M for nitrate and a sensitivity of 52 +/- 2 mV dec-1. The response time of the device is as low as 50 s, and the operational time of the system is ca. 85 s. We estimate the surface coverage of nitrate reductase on the gate surface to be ca. 1.2 x 10(-12) mol cm-2.
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PMID:An integrated relay/nitrate reductase field-effect transistor for the sensing of nitrate (NO3-). 1139 8

Rhodococcus sp. RB1 was able to thrive in media with up to 0.9 M NaCl or KCl and in the presence of high concentrations of nitrate (up to 0.9 M) and nitrite (up to 60 mM), but only under oxic conditions. An adaptation period was not required for salt tolerance, but a rapid extrusion of K+ and intake of Na+ was observed after addition of 0.5 M NaCl. Nitrate assimilation was limited by the carbon supply, but nitrite was not accumulated in the culture medium, even at nitrate concentrations as high as 0.8 M, thus suggesting that nitrite reduction does not limit nitrate assimilation. The presence of NaCl or KCl did not affect nitrate or nitrite uptake, which were completely inhibited by ammonium or glutamine. Rhodococcus sp. RB1 nitrate reductase had an apparent molecular mass of 142 kDa and used NADH and reduced bromophenol blue or viologens as electron donors, independently of the presence of salt. The enzyme was associated with an NADH-diaphorase activity and was induced by nitrate and repressed by ammonium or glutamine, thus showing typical biochemical and regulatory properties of bacterial assimilatory NADH-nitrate reductases. The enzyme was active in vitro in the presence of 3 M NaCl or KCI, but the maximal activity was observed at 0.5 M salt. Addition of 2 M NaCl increased the optimal temperature of the enzyme from 12 to 32 degrees C, but the optimal pH (10.3) was unaffected.
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PMID:Rhodococcus sp. RB1 grows in the presence of high nitrate and nitrite concentrations and assimilates nitrate in moderately saline environments. 1149 Oct 84

Nitrate reduction to N2O was investigated in batch cultures of Shewanella putrefaciens MR-1, MR-4, and MR-7. All three strains reduced nitrate to nitrite to N2O, and this reduction was coupled to growth, whereas ammonium accumulation was very low (0 to 1 micromol liter-1). All S. putrefaciens isolates were also capable of reducing nitrate aerobically; under anaerobic conditions, nitrite levels were three- to sixfold higher than those found under oxic conditions. Nitrate reductase activities (31 to 60 micromol of nitrite min-1 mg of protein-1) detected in intact cells of S. putrefaciens were equal to or higher than those seen in Escherichia coli LE 392. Km values for nitrate reduction ranged from 12 mM for MR-1 to 1.3 mM for MR-4 with benzyl viologen as an artifical electron donor. Nitrate and nitrite reductase activities in cell-free preparations were demonstrated in native gels by using reduced benzyl viologen. Detergent treatment of crude and membrane extracts suggested that the nitrate reductases of MR-1 and MR-4 are membrane bound. When the nitrate reductase in MR-1 was partially purified, three subunits (90, 70, and 55 kDa) were detected in denaturing gels. The nitrite reductase of MR-1 is also membrane bound and appeared as a 60-kDa band in sodium dodecyl sulfate-polyacrylamide gels after partial purification.
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PMID:Physiology and enzymology involved in denitrification by Shewanella putrefaciens. 1153 13

Studies on the diurnal variations of nitrate reductase (NR) activity during the life cycle of synchronized Chlorella sorokiniana cells grown with a 7:5 light-dark cycle showed that the NADH:NR activity, as well as the NR partial activities NADH:cytochrome c reductase and reduced methyl viologen:NR, closely paralleled the appearance and disappearance of NR protein as shown by sodium dodecyl sulfate gel electrophoresis and immunoblots. Results of pulse-labeling experiments with [35S]methionine further confirmed that diurnal variations of the enzyme activities can be entirely accounted for by the concomitant synthesis and degradation of the NR protein.
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PMID:Synthesis and degradation of nitrate reductase during the cell cycle of Chlorella sorokiniana. 1153 47


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