Competition between oxygen and nitrate respirations in continuous culture of Pseudomonas aeruginosa performing aerobic denitrification

Continuous culture of P. aeruginosa was conducted with nitrate-containing media under the dilution rates (D) of 0.026, 0.06, and 0.13/h and the dissolved oxygen concentrations (DO) of 0-2.2 mg/L. The bacterium performed simultaneous O(2) and nitrate respiration in all of the systems studied. For each D, the (apparent) cell yield from glucose (Y(X/S)) was lower at zero DO, but did not change substantially with non-zero DO. In non-zero DO systems, Y(X/S) increased with increasing D, and when fit with a model considering cell death, gave the following parameters: maximum cell yield Y(X/S) (m) = 0.49, maintenance coefficient M(S) = 0.029 (/h), and cell decay constant k(d) = 0.014/h. The same model failed to describe the behaviors of zero-DO systems, where neither glucose nor nitrate was limiting and the limiting factor(s) remained unknown. The cell yield from accepted electron (Y(X/e)) was however relatively constant in all systems, and the energy yield per electron accepted via denitrification was estimated at approximately 69% of that via O(2) respiration. A closer examination revealed that increasing DO enhanced O(2) respiration only at extremely low DO ( <0.05 mg/L), beyond which the increasing DO only slightly increased its weak inhibition on denitrification. While O(2) was the preferred electron acceptor, the fraction of electrons accepted via denitrification increased with increasing D.     

Microcalorimetric investigations of the host-parasite relationship between Biomphalaria glabrata and Schistosoma mansoni (author's transl)

1. The ratio of dry weight to wet weight of infected snails (5.5 +/- 2.2%) is significantly smaller than that of uninfected animals (8.3 +/- 1.0%). 2. The specific values of oxygen consumption and heat-production per dry weight are significantly higher for infected snails (2.41 +/- 0.44 microliter O2/h/mg; 12.63 +/- 1.24 muW/mg) than for uninfected ones (1.88 +/- 0.24 microliter O2/h/mg, 8.94 +/- 0.91 muW/mg). 3. The rate of oxygen consumption always suffices to explain the measured heat flows for aerobic catabolism of carbohydrates or fats and proteins, respectively. Neither for infected nor for uninfected snails is it necessary to assume an anaerobic energy metabolism. 4. Structures in the calorimetrically obtained heat production curves are discussed.     

Affinity chromatography of Ankistrodesmus braunii nitrate reductase using blue dextran-sepharose (author's transl)

Blue Dextran has been coupled covalently to Sepharose-4B to purify the enzymatic complex NAD(P)H-nitrate reductase (EC 1.6.6.2) from the green alga Ankistrodesmus braunii by affinity chromatography. The optimum conditions for the accomplishment of the chromatographic process have been determined. The adsorption of nitrate reductase on Blue Dextran Sepharose is optimum when a phosphate buffer of low ionic strength and pH 6.5-7.0 is used. Once the enzyme has been bound to Blue Dextran Sepharose, it can be specifically eluted by addition of NADH and FAD to the washing buffer. However, none of the nucleotides added separately is able to promote the elution of the enzyme from the column. The elution can be also achieved, but not specifically, by increasing the ionic strength of the buffer with KCl. These results have made possible a procedure for the purification of A. braunii nitrate reductase which led to electrophoretic homogeneity, with an overall yield of 70% and a specific activity of 49 units/mg of protein.