Low-temperature spectral and kinetic properties of cytochromes in Escherichia coli K-12 grown at lowered oxygen tension.
Escherichia coli K-12 was grown in batch culture in a medium containing succinate as carbon source, supplemented with casein hydrolysate, and with a rate of oxygen supply that resulted in dissolved O2 tension falling to 10% of saturation in the latter stages of growth. Cytochromes in such cells were qualitatively indistinguishable from those present in cells grown under conditions of vigorous aeration where dissolved O2 tensin remained greater than 80% saturation. Spectra recorded at 77 K and their fourth-order finite difference analyses revealed the absence of cytochrome b-558 and only low concentrations of cytochromes a1 and d(a2). At low temperatures, the reaction of cytochrome o with O2 in intact cells, grown under lowered O2 tension, proceeds through the same stage as observed previously in cells grown with vigorous aeration (Poole, R.K., Waring, A.J. and Chance, B. (1979) Biochem. J. 184, 379-389). However, much higher temperatures are required for comparable progress of the reaction in cells grown at lowered O2 tensions. AT 91 degrees C, the reaction with O2 involves ligand binding to give intermediate(s) with spectral characteristics similar to those of the reduced oxidase-CO complex. Temperatures of approx. -79 degrees C are required for the observation of biphasic kinetics and the attainment of an 'end point' in the reaction, features that are seen at temperatures below -98 degrees C in cells from vigorously-aerated cultures. At -32.5 degrees C, oxidation of cytochrome o is observed. The energy of activation for this  reaction at low temperatures is 29.9 kJ x mol-1. Binding with CO, in contrast to binding with O2, is characterized by high photolytic reversibility and appears to be less affected by the degree of aeration of cells during growth.
D003080:Cold Temperature; D003580:Cytochromes / Q000378:metabolism*; D004926:Escherichia coli / Q000254:growth & development / Q000378:metabolism*; D007700:Kinetics; D010100:Oxygen / Q000494:pharmacology*; D013057:Spectrum Analysis
D003580:Cytochromes; D010100:Oxygen