scholarly journals Nitrogenase of Klebsiella pneumoniae. Inhibition of acetylene reduction by magnesium ion explained by the formation of an inactive dimagnesium–adenosine triphosphate complex

1974 ◽  
Vol 139 (1) ◽  
pp. 211-214 ◽  
Author(s):  
Roger N. F. Thorneley ◽  
Keith R. Willison

Acetylene-reducing activity of purified nitrogenase from Klebsiella pneumoniae was studied over a range of ATP and Mg2+ concentrations at 15°C, pH7.8. Inhibition at Mg2+ concentrations of 2.5–30mm was due to the formation of the inactive complex, Mg2ATP. At higher Mg2+ concentrations an additional inhibitory effect was observed. The results were consistent with a MgATP complex being the active substrate with an apparent Km(MgATP)=0.4mm.

1981 ◽  
Vol 27 (1) ◽  
pp. 52-56 ◽  
Author(s):  
L. V. Wood ◽  
R. V. Klucas ◽  
R. C. Shearman

Turfs of 'Park' Kentucky bluegrass reestablished in the greenhouse and inoculated with Klebsiella pneumoniae (W6) showed significantly increased nitrogen fixation (acetylene reduction) compared with control turfs. Mean ethylene production rates per pot were 368 nmol h−1 for K. pneumoniae treated turfs, 55 nmol h−1 for heat-killed K. pneumoniae treated turfs, and 44 nmol h−1 for untreated turfs. Calculated lag periods before activity was observed were generally very short (less than 1 h).When 'Park' Kentucky bluegrass was grown from seed on soil-less medium of Turface, a fired aggregate clay, inoculation with K. pneumoniae (W6) resulted in 9 of 11 turfs showing nitrogenase activity (mean ethylene producion rate per pot was 195 nmol h−1). Only 3 of 11 turfs treated with heat-killed K. pneumoniae showed any activity and their mean rate of ethylene production (40 nmol h−1 per pot) was significantly lower than that for turfs treated with K. pneumoniae.Using the 'Park'–Turface soil-less model system it was shown that acetylene reducing activity was (i) root associated, (ii) generally highest at a depth of 1–4 cm below the surface, (iii) enhanced by washing excised roots, and (iv) inhibited by surface sterilization of excised roots. Klebsiella pneumoniae was recovered from Turface and roots showing acetylene reducing activity.


1981 ◽  
Vol 27 (8) ◽  
pp. 815-823 ◽  
Author(s):  
John D. Tjepkema ◽  
William Ormerod ◽  
John G. Torrey

Vesicle formation and acetylene reduction (nitrogenase activity) were observed when washed hyphae from cultures of Frankia sp. CpI1 were transferred to a nitrogen-free medium containing ethylenediaminetetraacetic acid and succinate. Succinate could be replaced by malate or fumarate, but not other carbon sources. Maximum acetylene reduction and vesicle numbers were observed at a pH of 6.0–6.5, at 25–30 °C, and at atmospheric [Formula: see text] or somewhat less (5–20 kPa). Addition of 1 mM NH4Cl almost completely inhibited vesicle formation and acetylene-reducing activity, but did not immediately inhibit such reducing activity by cultures with preexisting vesicles. Acetylene-reducing activity was never observed in the absence of vesicle formation.


1977 ◽  
Vol 23 (5) ◽  
pp. 567-572 ◽  
Author(s):  
T. Sekiguchi ◽  
A. Noguchi ◽  
Y. Nosoh

A sulfate-reducing bacterium, a strain of Desulfovibrio vulgaris, when lactate or pyruvate was supplied as an electron and energy source, reduced acetylene only in the presence of sulfate. Acetylene reduction started after a lag of 1 h. H2 which acts as an electron and energy source for sulfate reduction was unable to reduce acetylene even in the presence of sulfate. It was suggested, from the measurements of the amounts of adenine nucleotides of the bacterium incubated under various conditions, that the bacterium is able to reduce acetylene at a high ATP level or a high ATP/ADP ratio.


1989 ◽  
Vol 67 (2) ◽  
pp. 360-364 ◽  
Author(s):  
F. D. H. Macdowall ◽  
G. T. Kristjansson

Seedlings of three cultivars of alfalfa (Medicago sativa L.) were nodulated with two strains of rhizobia (Rhizobium meliloti) and grown with zero N nutrients at 25:20 °C for 6 weeks followed by growth at 10:7 °C for 2 weeks. Acetylene-reducing activity (ARA) was retarded by 10% acetylene to an extent dependent on time, cultivar, strain, and growth temperature. In the usual short term assay for nitrogenase by ARA, the inhibition was not sufficient to explain decreased apparent efficiency of nitrogenase in plants moved to the lower temperature. Inhibition of ARA was associated with correspondingly decreased respiration in cv. Drylander, but in cv. Apollo respiration was not affected. The differential loss of ARA relative to respiratory activity in a day of continuous treatment with 10% acetylene was very distinct and requires an explanation other than altered nodule resistance to diffusion of oxygen.


2004 ◽  
Vol 101 (2) ◽  
pp. 390-398 ◽  
Author(s):  
Takashi Kawano ◽  
Shuzo Oshita ◽  
Akira Takahashi ◽  
Yasuo Tsutsumi ◽  
Yoshinobu Tomiyama ◽  
...  

Background Sarcolemmal adenosine triphosphate-sensitive potassium (KATP) channels in the cardiovascular system may be involved in bupivacaine-induced cardiovascular toxicity. The authors investigated the effects of local anesthetics on the activity of reconstituted KATP channels encoded by inwardly rectifying potassium channel (Kir6.0) and sulfonylurea receptor (SUR) subunits. Methods The authors used an inside-out patch clamp configuration to investigate the effects of bupivacaine, levobupivacaine, and ropivacaine on the activity of reconstituted KATP channels expressed in COS-7 cells and containing wild-type, mutant, or chimeric SURs. Results Bupivacaine inhibited the activities of cardiac KATP channels (IC50 = 52 microm) stereoselectively (levobupivacaine, IC50 = 168 microm; ropivacaine, IC50 = 249 microm). Local anesthetics also inhibited the activities of channels formed by the truncated isoform of Kir6.2 (Kir6.2 delta C36) stereoselectively. Mutations in the cytosolic end of the second transmembrane domain of Kir6.2 markedly decreased both the local anesthetics' affinity and stereoselectivity. The local anesthetics blocked cardiac KATP channels with approximately eightfold higher potency than vascular KATP channels; the potency depended on the SUR subtype. The 42 amino acid residues at the C-terminal tail of SUR2A, but not SUR1 or SUR2B, enhanced the inhibitory effect of bupivacaine on the Kir6.0 subunit. Conclusions Inhibitory effects of local anesthetics on KATP channels in the cardiovascular system are (1) stereoselective: bupivacaine was more potent than levobupivacaine and ropivacaine; and (2) tissue specific: local anesthetics blocked cardiac KATP channels more potently than vascular KATP channels, via the intracellular pore mouth of the Kir6.0 subunit and the 42 amino acids at the C-terminal tail of the SUR2A subunit, respectively.


1984 ◽  
Vol 105 (1) ◽  
pp. 83-86 ◽  
Author(s):  
Donald L. Curry ◽  
Leslie L. Bennett

Abstract. Rat pancreas perfusions were performed using a perfusate with a fixed calcium concentration of 5 mEq/l and magnesium varying from 0 to 0.6 mEq/dl. Insulin secretion was stimulated by a constant glucose infusion of 300 mg/dl. This glucose concentration produces the typical biphasic insulin secretory response. We observed that in the absence of magnesium, somatostatin concentrations of 0.5 and 2.0 ng/ml were without effect on first phase insulin secretion. However, these same somatostatin levels produced 50% or more inhibition of insulin secretion in the presence of magnesium at 0.3 or 0.6 mEq/l. Similarly, in the absence of magnesium, somatostatin at 50 ng/ml failed to inhibit second phase insulin secretion, whereas this same somatostatin level produced about 50% inhibition of insulin secretion in the presence of magnesium at 0.3 mEq/l. Thus, altering perfusate magnesium concentrations without changing calcium is an important determinant of the degree of inhibition of secretion produced by somatostatin. In particular, in the absence of magnesium ion, somatostatin concentrations which would 'normally' produce 50% inhibition of secretion (ID50) are without effect. Therefore, magnesium ion is necessary for the full inhibitory effect of somatostatin to occur. These results suggest that inhibitors, as well as potentiators, of the insulin secretory process may act by altering intracellular/membrane calcium-magnesium ratios, but in opposite directions.


2003 ◽  
Vol 98 (1) ◽  
pp. 104-113 ◽  
Author(s):  
Ju-Tae Sohn ◽  
Paul A. Murray

Background The authors recently demonstrated that etomidate and ketamine attenuated endothelium-dependent pulmonary vasorelaxation mediated by nitric oxide and Ca -activated K + channels. In the current study, they tested the hypothesis that these intravenous anesthetics inhibit pulmonary vasorelaxation mediated by adenosine triphosphate-sensitive potassium (K + ATP ) channel activation. Methods Endothelium intact and denuded pulmonary arterial rings were suspended in organ chambers for isometric tension recording. The effects of etomidate (5 x 10(-6) and 5 x 10(-5) m) and ketamine (5 x 10(-5) and 10(-4) m) on vasorelaxation responses to lemakalim (K + ATP channel activator), prostacyclin, and papaverine were assessed in phenylephrine-precontracted rings. The effect of cyclooxygenase inhibition with indomethacin was assessed in some protocols. Results Etomidate (5 x 10(-6) m) only inhibited the vasorelaxant response to lemakalim in endothelium intact rings, whereas a higher concentration of etomidate (5 x 10(-5) m) inhibited relaxation in both intact and endothelium-denuded rings. Pretreatment with indomethacin abolished the endothelium-dependent attenuation of lemakalim-induced relaxation caused by etomidate. Ketamine (5 x 10(-5) and 10(-5) m) inhibited the relaxation response to lemakalim to the same extent in both endothelium-intact and -denuded rings, and this effect was not prevented by indomethacin pretreatment. Etomidate and ketamine had no effect on the relaxation responses to prostacyclin or papaverine. Conclusions These results indicate that etomidate, but not ketamine, attenuates the endothelium-dependent component of lemakalim-induced pulmonary vasorelaxation an inhibitory effect on the cyclooxygenase pathway. Both anesthetics inhibit K + ATP -mediated pulmonary vasorelaxation a direct effect on pulmonary vascular smooth muscle.


Author(s):  
O. R. Umeh ◽  
E. I. Chukwura ◽  
E. L. Okoye ◽  
E. M. Ibo ◽  
P. I. Egwuatu ◽  
...  

Medicinal plants are used by almost 80% of the world’s population for their basic health care because of their low cost and ease in availability. In the last few decades, many bacteria have continued to show increasing resistance against current antibiotics. Aim: In this study, phytochemical screening and antibacterial effects of conventional antibiotics, garlic and ginger on test isolates from fish pond water samples were evaluated between May-November, 2019. Methods: Standard methods for phytochemical screening and antibacterial analysis were employed. Results: The results showed that amongst the antibiotics used for susceptibility test, Amoxicilin (30 µg) was mostly resisted by all the bacterial isolates except Staphylococcus aureus, Vibrio cholerae, Vibrio parahaemolyticus and Salmonella typhi while erythromycin (10 µg) was unable to inhibit Bacillus subtilis. Ciprofloxacin (10 µg) and pefloxacin (10 µg) inhibited the growth of all the isolates except Pseudomonas aeruginosa. The test isolates showed variable susceptibility to the garlic and ginger extracts (ethanol, methanol and hot water). The extracts inhibited the isolates in descending order; ginger ethanol > ginger methanol > garlic methanol > ginger hot water > garlic ethanol > garlic hot water. Vibrio parahaemolyticus, Vibrio cholerae and Staphylococcus aureus showed little resistant to the extracts while these extracts showed better activity on Klebsiella pneumoniae and Proteus mirabilis. Synergistic effect of garlic and ginger (500mg/ml) inhibited the growth of all the isolates with ethanol extracts having the highest zone diameter (29 mm) on Klebsiella pneumoniae and Proteus mirabilis while hot water extracts had the least zone of inhibition (18 mm) on Acinetobacter calcoaceticus and Vibrio parahaemolyticus. The minimum inhibitory and bactericidal concentration for ethanol, methanol and hot water extracts ranged from 31.25mg/ml to 62.5mg/ml and 62.5mg/ml to 125mg/ml respectively. Conclusion: The outcomes of susceptibility experiment depicted that ethanol and methanol extracts of garlic and ginger (each alone and in combination) showed more inhibitory effect than aqueous extracts and also the combination of ethanol, methanol and aqueous extracts resulted in inhibitory effect greater than each of the extracts when used singly. The use of ginger and garlic for control of fish pathogens appears to be justified.


1983 ◽  
Vol 61 (9) ◽  
pp. 2405-2409 ◽  
Author(s):  
F. D. H. Macdowall

Seedlings of Medicago sativa L. cv. Algonquin were grown in vermiculite and nodulated by Rhizobium meliloti strain 102F70 at two lower levels of N, until flowering when the tops were cut off to leave about 10% shoot stubble. Residual shoot dry matter immediately resumed first-order growth and maintained it throughout regrowth to second flowering. The rate constants of shoot regrowth were 34% lower (at 15 mM NO3−), 25% lower (at 1.5 mM NO3− symbiotically), or 220% higher (at zero NO3− symbiotically) than the values for 1 to 4-week-old seedlings, which indicated a radical change in physiology. Root dry matter resumed exponential growth after a 7-day recession and its recovery and yields were independent of N nutrition. The most pronounced minima occurred in the acetylene-reducing activity of nitrogenase, the kinetics of which paralleled root dry matter except that its redevelopment stopped after two-thirds of the regrowth time. The rate coefficient for the redevelopment of nitrogenase activity equalled that for its development during the seedling stage, which suggested unchanged limitations on that process until its redevelopment stopped.


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