Respiratory behaviour of a Zymomonas mobilis adhB::kan(r) mutant supports the hypothesis of two alcohol dehydrogenase isoenzymes catalysing opposite reactions.

AbstractPerturbation of the aerobic steady-state in a chemostat culture of the ethanol-producing bacterium Zymomonas mobilis with a small pulse of ethanol causes a burst of ethanol oxidation, although the reactant ratio of the alcohol dehydrogenase (ADH) reaction ([NADH][acetaldehyde][H+])/([ethanol][NAD+]) remains above the Keq value. Simultaneous catalysis of ethanol synthesis and oxidation by the two ADH isoenzymes, residing in different redox microenvironments, has been proposed previously. In the present study, this hypothesis is verified by construction of an ADH-deficient strain and by demonstration that it lacks the oxidative burst in response to perturbation of its aerobic steady-s…

Ethanol cycle in an ethanologenic bacterium

AbstractA novel redox cycle is suggested, performing interconversion between acetaldehyde and ethanol in aerobically growing ethanologenic bacterium Zymomonas mobilis. It is formed by the two alcohol dehydrogenase (ADH) isoenzymes simultaneously catalyzing opposite reactions. ADH I is catalyzing acetaldehyde reduction. The local reactant ratio at its active site probably is shifted towards ethanol synthesis due to direct channeling of NADH from glycolysis. ADH II is oxidizing ethanol. The net result of the cycle operation is NADH shuttling from glycolysis to the membrane respiratory chain, and ensuring flexible distribution of reducing equivalents between the ADH reaction and respiration.

Erratum to “electron transport chain in aerobically cultivatedZymomonas mobilis” [FEMS microbiol. lett. 143 (1996) 185–189]

Cultivation ofZymomonas mobilis 113 S at Different Mixing Regimes and their Influence on the Levan Formation

The Zymomonas mobilis I 13 S strain was cultivated in a bioreactor with a working volume of 1.4 I at different stirring regimes in a 15% initial sucrose medium The levan obtained in the fermentation process was analyzed by gel filtration. Because the sucrose/biomass ratio in the fermentation broth decreased to below 300 g/g, the insufficient concentration of sucrose might have decreased the concentration of levan. Besides the growth characteristics of the population, the mixing intensity and flow structure were also found to influence the molecular mass of levan. At 600 rpm, the microorganisms produced levan with a molecular mass lower than at 300 rpm. The stirring of a fermentation broth w…

The effect of osmo-induced stress on product formation by Zymomonas mobilis on sucrose

The intensification of biosynthesis of fructooligosaccharides in the presence of high salt concentrations was observed during sucrose (10%) fermentation by Zymomonas mobilis 113S. A 0.6 M NaCl concentration led to an increase of oligosaccharide productivity by 3.5-fold. Sorbitol formation was increased in the presence of 0.16 M NaCl and was inhibited at highest salt concentrations. In a medium with high (65%, w/w) sucrose content the salts gave inhibitory effects on fructooligosaccharide production by lyophilised Z. mobilis cells. Influence of salts on gluconic acid and sorbitol formation under these conditions was studied. The ratio of oligosaccharides and gluconic acid productivity (Qolig…

The effect of mixing on glucose fermentation by Zymomonas mobilis continuous culture

Abstract The influence of mixing (in the range from 300 to 1100 rpm) on the performance of Zymomonas mobilis anaerobic continuous culture was studied. Biomass yield and ethanol productivity were improved at higher stirring intensities, in parallel with a decrease in byproduct synthesis. A mixing-dependent formation of minor amounts of fructose was first demonstrated in Z. mobilis anaerobic culture on glucose. The rate of fructose synthesis and the specific activity of the key enzyme in the pathway to fructose, phosphoglucose isomerase (EC 5.3.1.9), had a maximum at 700 rpm.

Electron transport chain in aerobically cultivated Zymomonas mobilis

Respiratory chain composition and energy coupling in cytoplasmic membrane of Zymomonas mobilis was shown to depend on culture aeration. Aerobically grown cells contained mainly the non-energy-generating NADH dehydrogenase with kM for NADH 58 mM. In anaerobically cultivated bacteria, the energy-coupling NADH dehydrogenase complex with km for NADH 7 mM predominated. In aerobically cultivated Z. mobilis, CoQ content and absorption peaks of cytochromes at 554–556 nm and 525–528 nm were significantly increased. Energy-coupling site I, linked to the NADH:CoQ oxidoreductase complex, could be eliminated under sulfate-deficient cultivation conditions. For anaerobically grown cells this resulted in l…

Cyanide inhibits respiration yet stimulates aerobic growth of Zymomonas mobilis

Potassium cyanide at submillimolar concentrations (20-500 microM) inhibited the high respiration rates of aerobic cultures of Zymomonas mobilis but, remarkably, stimulated culture growth. In batch culture, after an extended lag phase, exponential growth persisted longer, resulting in higher biomass densities. In aerobic chemostat cultures, elevated biomass concentration was observed in the presence of cyanide. This growth stimulation effect is attributed to decreased production of the inhibitory metabolite acetaldehyde at lowered respiration rates, when more reducing equivalents are channelled to alcohol dehydrogenase. Growth in the presence of cyanide did not alter the membrane cytochrome …

The paradoxical cyanide-stimulated respiration of Zymomonas mobilis: cyanide sensitivity of alcohol dehydrogenase (ADH II)

The respiratory inhibitor cyanide stimulates growth of the ethanologenic bacteriumZymomonas mobilis, perhaps by diverting reducing equivalents from respiration to ethanol synthesis, thereby minimizing accumulation of toxic acetaldehyde. This study sought to identify cyanide-sensitive components of respiration. In aerobically grown, permeabilizedZ. mobiliscells, addition of 200 μM cyanide caused gradual inhibition of ADH II, the iron-containing alcohol dehydrogenase isoenzyme, which, in aerobic cultures, might be oxidizing ethanol and supplying NADH to the respiratory chain. In membrane preparations, NADH oxidase was inhibited more rapidly, but to a lesser extent, than ADH II. The time-cours…

Influence of Micromixing on Microorganisms and Products

The influence of mixing intensity as well as physical and chemical parameters on the cells of different microorganisms and the biosynthesis process is examined in this paper. Some reactions of cells effecting mixing intensity are described, such as retarded biomass growth, changes in aggregation and mutual arrangement of cells, morphological changes of cells and decreasing of biological activity, caused by an increased intensity of turbulence (turbohypobiosis). Several methods for investigating the local energy in reactors are compared. It is concluded that conventional methods of hydrodynamic analysis do not always allow valid results for the optimization of the mixing regime to be obtaine…

The effect of probiotics on the genotoxicity of furazolidone.

Antigenotoxic activity of probiotic bacteria against furazolidone was studied using the short-term bacterial assay SOS chromotest, with Escherichia coli PQ37 as the test organism. The supernatants from probiotic and furazolidone co-incubation exhibited rather strong suppression on SOS induction produced by furazolidone on E. coli PQ 37 (sfiA: lacZ). Genotoxicity inhibition was found for all strains of the examined bacteria belonging to three genera. The highest genotoxicity inhibition was detected for Bifidobacterium lactis Bb-12 (92.0%) and for Lactobacillus acidophilus T20 (81.9%).