Thiamine pyrophosphate stimulates acetone activation by Desulfococcus biacutus as monitored by a fluorogenic ATP analogue.
Acetone can be degraded by aerobic and anaerobic microorganisms. Studies with the strictly anaerobic sulfate-reducing bacterium Desulfococcus biacutus indicate that acetone degradation by these bacteria starts with an ATP-dependent carbonylation reaction leading to acetoacetaldehyde as the first reaction product. The reaction represents the second example of a carbonylation reaction in the biochemistry of strictly anaerobic bacteria, but the exact mechanism and dependence on cofactors are still unclear. Here, we use a novel fluorogenic ATP analogue to investigate its mechanism. We find that thiamine pyrophosphate is a cofactor of this ATP-dependent reaction. The products of ATP cleavage are AMP and pyrophosphate, providing first insights into the reaction mechanism by indicating that the reaction proceeds without intermediate formation of acetone enol phosphate.
Department of Biology and ‡Department of Chemistry, Konstanz Research School Chemical Biology, University of Konstanz , Universitätsstr. 10, 78457 Konstanz, Germany.
Olga B
OB
Gutiérrez Acosta
Norman
N
Hardt
Stephan M
SM
Hacker
Tobias
T
Strittmatter
Bernhard
B
Schink
Andreas
A
Marx
D000096:Acetone / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000737:chemistry*; D020564:Deltaproteobacteria / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry*; D013835:Thiamine Pyrophosphate / Q000031:analogs & derivatives* / Q000378:metabolism*
D005456:Fluorescent Dyes; D000096:Acetone; D000255:Adenosine Triphosphate; D013835:Thiamine Pyrophosphate