Respiratory differences associated with culture aeration in Azotobacter vinelandii

1989 ◽  
Vol 35 (10) ◽  
pp. 918-924 ◽  
Author(s):  
Jay B. Peterson

Respiratory oxygen uptake of nitrogen-fixing Azotobacter vinelandii cells was altered by culturing at different levels of culture agitation (aeration). Cells were grown at low agitation or moderate agitation and with three different carbon substrates. The low-agitation cultures had much lower dissolved-oxygen concentrations than moderate-agitation cultures at the stage of growth at which they were studied. The respiration systems of cells from the moderate-agitation cultures had lower apparent affinities (higher Ks (O2) values) for oxygen than cells grown at low agitation. The higher Ks (O2)) values were dependent on the presence of Ca2+ and (or) Mg2+ in the medium. In low-agitation cultures, the oxygen concentrations were below the Ks (O2 values and the respiration rates in the cultures were therefore well below the maximal respiration (Vmax) rates. The oxygen concentrations in moderate-agitation cultures were above the Ks (O2) values and the culture respiration rates were much higher. The culture oxygen concentration relative to the Ks (O2) had a much greater effect on culture respiration rate than did the Vmax. It is proposed that changes in the respiration system resulting from culture agitation (aeration) reflect an "oxygen-sensing mechanism" that regulates respiration. This would provide at least a partial explanation for the increased respiration rates with increased culture oxygen concentration in A. vinelandii.Key words: Azotobacter, oxygen, respiration, nitrogen fixation.

1991 ◽  
Vol 37 (4) ◽  
pp. 321-325 ◽  
Author(s):  
Jay B. Peterson

The effects of three low oxygen concentrations on nitrogenase activity, total oxygen uptake, and respiratory parameters (Vmax and Ks(O2) of N2-grown Azotobacter vinelandii were studied in acetylene reduction assays during a 2-h incubation. The cell suspensions were taken from cultures grown at low aeration. Total oxygen uptake was higher with each increment in oxygen concentration. The highest oxygen concentration was initially supraoptimal for nitrogenase activity. The Ks(O2) values, representing the apparent affinity of the respiration system for oxygen, increased during the incubation of cells at the highest oxygen concentration. The Ks(O2) values at the two lower oxygen concentrations decreased and were very similar. A small effect of oxygen on the Vmax was observed. These results show that the metabolism determining the apparent affinity of the system for oxygen responds to the oxygen concentrations. Furthermore, this metabolism did not substantially increase the Ks(O2) unless the oxygen concentration was high enough to inhibit nitrogenase activity, indicating that the two processes may be linked. Key words: Azotobacter, oxygen regulation, nitrogen fixation.


1963 ◽  
Vol 204 (1) ◽  
pp. 5-8 ◽  
Author(s):  
Kalman Greenspan ◽  
Paul F. Cranefield

The rate of oxygen uptake of quiescent Purkinje fibers of the dog's heart was determined using a flow respirometer and oxygen polarography. At ambient oxygen concentrations of 60% or higher the rate of uptake was 0.739 mm3/mg wet weight per hr at 35 C. The temperature coefficient over the range 25–35° was 2.3. The uptake was independent of the ambient oxygen concentration at oxygen concentrations equal to or greater than 60% of an atmosphere. In lower oxygen concentrations the rate of uptake was found to be depressed. The depression of uptake in the lower oxygen tensions is probably the result of diffusion limitation; it may, however, reflect dependence of resting uptake on oxygen concentration.


1989 ◽  
Vol 29 (5) ◽  
pp. 741 ◽  
Author(s):  
FC Miller ◽  
ER Harper ◽  
BJ Macauley

Temperature and oxygen concentration in Phase I composting stacks were investigated in the field. that both determine and are consequences of biological Investigations focused on various physical factors activity. Data indicate that oxygen concentrations in Phase I stacks are affected by convection, gaseous diffusion and utilisation rates, but that these rates vary significantly spatially and temporally. When stack temperatures exceed 60�C, biological rates of activity, and therefore oxygen uptake, decrease, allowing oxygen to penetrate well into the centres of stacks. While natural convection is commonly used to explain stack oxygenation, this is an over simplification that does not adequately describe stack oxygen concentrations.


1968 ◽  
Vol 14 (1) ◽  
pp. 33-38 ◽  
Author(s):  
M. C. Mahl ◽  
P. W. Wilson

A cell-free system which permits nitrogen fixation by extracts of Klebsiella pneumoniae M5al (formerly Aerobacter aerogenes) has been developed. It is, essentially, that system described by Bulen and associates for Azotobacter vinelandii, utilizing ATP as a source of energy and dithionite as a source of electrons. The Michaelis constant for fixation has been estimated to be 0.12 atm. The extracts possessed an ATP-dependent hydrogen evolving system. Hydrogen evolution from these extracts was less under nitrogen than under helium in the presence of ATP. Nitrogen gas appears to be the inducer of nitrogen fixation. In the absence of N2, no induction of nitrogenase occurs. Nitrogenase is absent in cells grown on NH4+-N. There is a lag of about 13 h after the introduction of N2 gas into a culture which has depleted its supply of NH4+-N before nitrogenase can be detected. For reasons discussed in the text, this conclusion must be regarded as tentative at this time. Ammonium ion appears to prevent the synthesis of new molecules of nitrogenase without affecting the activity of those already formed.


Parasitology ◽  
1974 ◽  
Vol 68 (2) ◽  
pp. 271-284 ◽  
Author(s):  
A. E. Rumpus ◽  
C. R. Kennedy

The respiration rates of individual Gammarus pulex infected by larval Pomphorhynchus laevis were investigated with particular reference to the stage of development of the host and parasite and to the water temperature. At 20°C the oxygen consumption of Gammarus of all sizes was reduced by an average of 19·3 % by the presence of cystacanths of the parasite, but was unaffected by the presence of acanthellae. It is considered that the small size of this larval stage, in relation to that of its host, is responsible for the failure to detect an effect. Multiple infections did not exert any greater effect upon host respiration than single cystacanths, nor did it appear that the parasite had different effects upon hosts of different sexes. At 10°C no significant differences were observed between the respiration rates of infected and uninfected gammarids. The parasite was probably still depressing the host respiration rate at this temperature, but the oxygen uptake of G. pulex is so low that the differences between infected and uninfected individuals were too small to be detected. The parasite has a direct effect upon the physiological processes of the host, but neither the mechanism of this nor the reasons for the different effects found in different host-parasite systems are yet understood. Despite the pronounced effect of P. laevis on respiration of individual hosts, its effect upon the oxygen consumption of a natural host population is small since only a small proportion of the population carries infections and water temperatures remain below 10°C for over half the year.


Author(s):  
Yulia V Bertsova ◽  
Marina V Serebryakova ◽  
Alexander A Baykov ◽  
Alexander V Bogachev

Abstract Azotobacter vinelandii, the model microbe in nitrogen fixation studies, uses the ferredoxin:NAD+-oxidoreductase Rnf to regenerate ferredoxin (flavodoxin) acting as an electron donor for nitrogenase. However, the relative contribution of Rnf into nitrogenase functioning is unknown because this bacterium contains another ferredoxin reductase, FixABCX. Furthermore, Rnf is flavinylated in the cell, but the importance and pathway of this modification reaction also remain largely unknown. We have constructed A. vinelandii cells with impaired activities of FixABCX and/or putative flavin transferase ApbE. The ApbE-deficient mutant could not produce covalently flavinylated membrane proteins and demonstrated a markedly decreased flavodoxin:NAD+ oxidoreductase activity and significant growth defect under diazotrophic conditions. The double ΔFix/ΔApbE mutation abolished the flavodoxin:NAD+ oxidoreductase activity and the ability of A. vinelandii to grow in the absence of fixed nitrogen source. ApbE flavinylated a truncated RnfG subunit of Rnf1 by forming a phosphoester bond between FMN and a threonine residue. These findings indicate that Rnf (presumably its Rnf1 form) is the major ferredoxin-reducing enzyme in the nitrogen fixation system and that the activity of Rnf depends on its covalent flavinylation by the flavin transferase ApbE.


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