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首页> 外文期刊>Journal of bacteriology >Glucose Metabolism and Acetate Switch in Archaea: the Enzymes in Haloferax volcanii
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Glucose Metabolism and Acetate Switch in Archaea: the Enzymes in Haloferax volcanii

机译：古代葡萄糖新陈代谢和醋酸醋酸酯开关：卤代克劳克斯火山的酶

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The halophilic archaeon Haloferax volcanii has been proposed to degrade glucose via the semiphosphorylative Entner-Doudoroff (spED) pathway. Following our previous studies on key enzymes of this pathway, we now focus on the characterization of enzymes involved in 3-phosphoglycerate conversion to pyruvate, in anaplerosis, and in acetyl coenzyme A (acetyl-CoA) formation from pyruvate. These enzymes include phosphoglycerate mutase, enolase, pyruvate kinase, phosphoenolpyruvate carboxylase, and pyruvate-ferredoxin oxidoreductase. The essential function of these enzymes were shown by transcript analyses and growth experiments with respective deletion mutants. Furthermore, we show that H. volcanii—during aerobic growth on glucose—excreted significant amounts of acetate, which was consumed in the stationary phase (acetate switch). The enzyme catalyzing the conversion of acetyl-CoA to acetate as part of the acetate overflow mechanism, an ADP-forming acetyl-CoA synthetase (ACD), was characterized. The functional involvement of ACD in acetate formation and of AMP-forming acetyl-CoA synthetases (ACSs) in activation of excreted acetate was proven by using respective deletion mutants. Together, the data provide a comprehensive analysis of enzymes of the spED pathway and of anaplerosis and report the first genetic evidence of the functional involvement of enzymes of the acetate switch in archaea. IMPORTANCE In this work, we provide a comprehensive analysis of glucose degradation via the semiphosphorylative Entner-Doudoroff pathway in the haloarchaeal model organism Haloferax volcanii. The study includes transcriptional analyses, growth experiments with deletion mutants. and characterization of all enzymes involved in the conversion of 3-phosphoglycerate to acetyl coenzyme A (acetyl-CoA) and in anaplerosis. Phylogenetic analyses of several enzymes indicate various lateral gene transfer events from bacteria to haloarchaea. Furthermore, we analyzed the key players involved in the acetate switch, i.e., in the formation (overflow) and subsequent consumption of acetate during aerobic growth on glucose. Together, the data provide novel aspects of glucose degradation, anaplerosis, and acetate switch in H. volcanii and thus expand our understanding of the unusual sugar metabolism in archaea.
机译：已经提出了嗜盐古氏卤代洛克斯型Volcanii以通过半磷酸化的Entner-Doudoroff（Sped）途径来降解葡萄糖。遵循我们之前对该途径的关键酶的研究，我们现在关注参与3-磷酸糖转化为丙酮酸的酶的酶，在丙酮中，乙酰辅酶A（乙酰-CoA）从丙酮酸形成。这些酶包括磷酸性蛋白酶，烯醇酶，丙酮酸激酶，磷酸丙酮酸羧酸盐和丙酮酸 - 富勒沙霉蛋白氧化酶。通过转录物分析和生长实验显示这些酶的基本函数，其缺失突变体。此外，我们表明H. Volcanii期间的葡萄糖生长在葡萄糖 - 排泄的大量醋酸盐上，其在固定相（醋酸开关）中消耗。表征催化乙酰-COA转化为乙酸酯的乙酸乙烯酯，其特征在于乙酸盐溢流机构的一部分。通过使用各自的缺失突变体证明，通过各自的缺失突变体证明ACD在乙酸盐形成和AMP形成乙酰-COA合成酶（ACSS）中的功能介断。在一起，数据可以全面分析Sped途径和AnaPlerosis的酶，并报告乙酸酸酶酶功能参与古痤疮的第一个遗传证据。重要性在这项工作中，我们通过HaloAthaeal模型生物中的半磷酸化Entner-Doudoroff途径进行全面分析葡萄糖降解卤素模型生物Haloferax Volcanii。该研究包括转录分析，缺失突变体的生长实验。并表征涉及3-磷酸糖至乙酰辅酶A（乙酰-CoA）和Anplerosis的转化率。几种酶的系统发育分析表明，来自细菌到HaloAthaea的各种外侧基因转移事件。此外，我们分析了醋酸酯开关中涉及的关键球员，即在葡萄糖的有氧生长期间的醋酸盐和随后的醋酸盐消耗。这些数据在一起提供了葡萄糖降解，Anpererosis和H. Volcanii的醋酸纤维切换的新颖方面，从而扩大了我们对古亚群岛异常糖代谢的理解。

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