Beschreibung:
<jats:p>The thermophilic methanogen<jats:italic>Methanosaeta thermophila</jats:italic>uses acetate as sole substrate for methanogenesis. It was proposed that the acetate activation reaction that is needed to feed acetate into the methanogenic pathway requires the hydrolysis of two ATP, whereas the acetate activation reaction in<jats:italic>Methanosarcina sp.</jats:italic>is known to require only one ATP. As these organisms live at the thermodynamic limit that sustains life, the acetate activation reaction in<jats:italic>Mt. thermophila</jats:italic>seems too costly and was thus reevaluated. It was found that of the putative acetate activation enzymes one gene encoding an AMP-forming acetyl-CoA synthetase was highly expressed. The corresponding enzyme was purified and characterized in detail. It catalyzed the ATP-dependent formation of acetyl-CoA, AMP, and pyrophosphate<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M1"><mml:mrow><mml:mrow><mml:mo stretchy="false">(</mml:mo><mml:mrow><mml:msub><mml:mrow><mml:mtext>PP</mml:mtext></mml:mrow><mml:mtext>i</mml:mtext></mml:msub></mml:mrow><mml:mo stretchy="false">)</mml:mo></mml:mrow></mml:mrow></mml:math>and was only moderately inhibited by<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M2"><mml:mrow><mml:msub><mml:mrow><mml:mtext>PP</mml:mtext></mml:mrow><mml:mtext>i</mml:mtext></mml:msub></mml:mrow></mml:math>. The breakdown of<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M3"><mml:mrow><mml:msub><mml:mrow><mml:mtext>PP</mml:mtext></mml:mrow><mml:mtext>i</mml:mtext></mml:msub></mml:mrow></mml:math>was performed by a soluble pyrophosphatase. This enzyme was also purified and characterized. The pyrophosphatase hydrolyzed the major part of<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M4"><mml:mrow><mml:msub><mml:mrow><mml:mtext>PP</mml:mtext></mml:mrow><mml:mtext>i</mml:mtext></mml:msub></mml:mrow></mml:math>(<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M5"><mml:mrow><mml:msub><mml:mrow><mml:mi>K</mml:mi></mml:mrow><mml:mrow><mml:mi>M</mml:mi></mml:mrow></mml:msub><mml:mo>=</mml:mo><mml:mn>0.27</mml:mn><mml:mo>±</mml:mo><mml:mn>0.05</mml:mn></mml:mrow></mml:math> mM) that was produced in the acetate activation reaction. Activity was not inhibited by nucleotides or<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M6"><mml:mrow><mml:msub><mml:mrow><mml:mtext>PP</mml:mtext></mml:mrow><mml:mtext>i</mml:mtext></mml:msub></mml:mrow></mml:math>. However, it cannot be excluded that other<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M7"><mml:mrow><mml:msub><mml:mrow><mml:mtext>PP</mml:mtext></mml:mrow><mml:mtext>i</mml:mtext></mml:msub></mml:mrow></mml:math>-dependent enzymes take advantage of the remaining<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M8"><mml:mrow><mml:msub><mml:mrow><mml:mtext>PP</mml:mtext></mml:mrow><mml:mtext>i</mml:mtext></mml:msub></mml:mrow></mml:math>and contribute to the energy balance of the cell.</jats:p>