Cytochrome bd oxidase, oxidative stress, and dioxygen tolerance of the strictly anaerobic bacterium Moorella thermoacetica.
The gram-positive, thermophilic, acetogenic bacterium Moorella thermoacetica can reduce CO2 to acetate via the Wood-Ljungdahl (acetyl coenzyme A synthesis) pathway. This report demonstrates that, despite its classification as a strict anaerobe, M. thermoacetica contains a membrane-bound cytochrome bd oxidase that can catalyze reduction of low levels of dioxygen. Whole-cell suspensions of M. thermoacetica had significant endogenous O2 uptake activity, and this activity was increased in the presence of methanol or CO, which are substrates in the Wood-Ljungdahl pathway. Cyanide and azide strongly (approximately 70%) inhibited both the endogenous and CO/methanol-dependent O2 uptake. UV-visible light absorption and electron paramagnetic resonance spectra of n-dodecyl-beta-maltoside extracts of M. thermoacetica membranes showed the presence of a cytochrome bd oxidase complex containing cytochrome b561, cytochrome b595, and cytochrome d (chlorin). Subunits I and II of the bd oxidase were identified by N-terminal amino acid sequencing. The M. thermoacetica cytochrome bd oxidase exhibited cyanide-sensitive quinol oxidase activity. The M. thermoacetica cytochrome bd (cyd) operon consists of four genes, encoding subunits I and II along with two ABC-type transporter proteins, homologs of which in other bacteria are required for assembly of the bd complex. The level of this cyd operon transcript was significantly increased when M. thermoacetica was grown in the absence of added reducing agent (cysteine + H2S). Expression of a 35-kDa cytosolic protein, identified as a cysteine synthase (CysK), was also induced by the nonreducing growth conditions. The combined evidence indicates that cytochrome bd oxidase and cysteine synthase protect against oxidative stress and contribute to the limited dioxygen tolerance of M. thermoacetica.
Das
Amaresh
A
Department of Chemistry and Molecular Biology, University of Georgia, Athens, GA 30602-2556, USA.
Silaghi-Dumitrescu
Radu
R
Ljungdahl
Lars G
LG
Kurtz
Donald M
DM
D000595:Amino Acid Sequence; D001421:Bacteria, Anaerobic / Q000235:genetics* / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D003545:Cysteine / Q000378:metabolism; D003547:Cysteine Synthase / Q000235:genetics / Q000378:metabolism; D003580:Cytochromes / Q000235:genetics* / Q000378:metabolism; D045222:Electron Transport Chain Complex Proteins / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D006418:Heme / Q000378:metabolism; D008969:Molecular Sequence Data; D009876:Operon / Q000502:physiology; D018384:Oxidative Stress / Q000235:genetics; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D010100:Oxygen / Q000493:pharmacokinetics
D003580:Cytochromes; D045222:Electron Transport Chain Complex Proteins; D006418:Heme; D010088:Oxidoreductases; C020094:duroquinol oxidase; D003547:Cysteine Synthase; D003545:Cysteine; D010100:Oxygen
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