Thiamine Pyrophosphate Stimulates Acetone Activation by Desulfococcus biacutus As Monitored by a Fluorogenic ATP Analogue
Thiamine Pyrophosphate Stimulates Acetone Activation by Desulfococcus biacutus As Monitored by a Fluorogenic ATP Analogue
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DOI：
10.1021/cb500152y
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发表时间：
2014-06-01
影响因子：
4
通讯作者：
Marx, Andreas
中科院分区：
文献类型：
--
作者：
Acosta, Olga B. Gutierrez;Hardt, Norman;Marx, Andreas
丙酮可以被好氧和厌氧微生物降解。严格厌氧硫酸盐还原细菌双尖脱硫球菌的研究表明，丙酮降解这些细菌开始与ATP依赖的羰基化反应，导致乙酰乙醛作为第一反应产物。该反应代表了严格厌氧细菌生物化学中羰基化反应的第二个例子，但确切的机制和对辅因子的依赖性仍然不清楚。在这里，我们使用一种新的荧光ATP类似物来研究其机制。我们发现焦磷酸硫胺素是这种ATP依赖性反应的辅因子。ATP裂解的产物是AMP和焦磷酸盐，通过表明反应在没有丙酮烯醇磷酸的中间体形成的情况下进行，提供了对反应机理的第一次了解。
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.