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反硝化嗜环脂菌BC菌株对硝酸盐依赖型丙酮降解的蛋白质组学分析

Proteomic analysis of nitrate-dependent acetone degradation by Alicycliphilus denitrificans strain BC.

作者信息

Oosterkamp Margreet J, Boeren Sjef, Atashgahi Siavash, Plugge Caroline M, Schaap Peter J, Stams Alfons J M

机构信息

Laboratory of Microbiology, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, the Netherlands.

Laboratory of Biochemistry, Wageningen University, Dreijenplein 3, 6703 HA Wageningen, the Netherlands.

出版信息

FEMS Microbiol Lett. 2015 Jun;362(11). doi: 10.1093/femsle/fnv080. Epub 2015 May 14.

Abstract

Alicycliphilus denitrificans strain BC grows anaerobically on acetone with nitrate as electron acceptor. Comparative proteomics of cultures of A. denitrificans strain BC grown on either acetone or acetate with nitrate was performed to study the enzymes involved in the acetone degradation pathway. In the proposed acetone degradation pathway, an acetone carboxylase converts acetone to acetoacetate, an AMP-dependent synthetase/ligase converts acetoacetate to acetoacetyl-CoA, and an acetyl-CoA acetyltransferase cleaves acetoacetyl-CoA to two acetyl-CoA. We also found a putative aldehyde dehydrogenase associated with acetone degradation. This enzyme functioned as a β-hydroxybutyrate dehydrogenase catalyzing the conversion of surplus acetoacetate to β-hydroxybutyrate that may be converted to the energy and carbon storage compound, poly-β-hydroxybutyrate. Accordingly, we confirmed the formation of poly-β-hydroxybutyrate in acetone-grown cells of strain BC. Our findings provide insight in nitrate-dependent acetone degradation that is activated by carboxylation of acetone. This will aid studies of similar pathways found in other microorganisms degrading acetone with nitrate or sulfate as electron acceptor.

摘要

反硝化脂环酸芽孢杆菌BC菌株在以硝酸盐作为电子受体的条件下,可利用丙酮进行厌氧生长。对反硝化脂环酸芽孢杆菌BC菌株在以丙酮或乙酸盐为底物、硝酸盐为电子受体的培养物进行了比较蛋白质组学研究,以探究参与丙酮降解途径的酶。在所提出的丙酮降解途径中,丙酮羧化酶将丙酮转化为乙酰乙酸,AMP依赖的合成酶/连接酶将乙酰乙酸转化为乙酰乙酰辅酶A,乙酰辅酶A乙酰基转移酶将乙酰乙酰辅酶A裂解为两个乙酰辅酶A。我们还发现了一种与丙酮降解相关的假定醛脱氢酶。该酶作为β-羟基丁酸脱氢酶发挥作用,催化多余的乙酰乙酸转化为β-羟基丁酸,β-羟基丁酸可转化为能量和碳储存化合物聚-β-羟基丁酸。因此,我们证实了BC菌株在以丙酮为底物生长的细胞中形成了聚-β-羟基丁酸。我们的研究结果为通过丙酮羧化激活的依赖硝酸盐的丙酮降解提供了见解。这将有助于研究其他以硝酸盐或硫酸盐作为电子受体降解丙酮的微生物中发现的类似途径。

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