Insights into the Substrate Specificity of Archaeal Entner–Doudoroff Aldolases: The Structures of <i>Picrophilus torridus</i> 2-Keto-3-deoxygluconate Aldolase and <i>Sulfolobus solfataricus</i> 2-Keto-3-deoxy-6-phosphogluconate Aldolase in Complex with 2-Keto-3-deoxy-6-phosphogluconate

Zaĭtsev, V. N.; Johnsen, Ulrike; Reher, Matthias; Ortjohann, Marius; Taylor, G.L.; Danson, Michael J.; Schönheit, Peter; Crennell, S.J.

doi:10.1021/acs.biochem.8b00535

Cited by 6 publications

(2 citation statements)

References 25 publications

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“…In particular, sugar metabolism has been studied in detail (1,2). It was found that anaerobic and microaerophilic hyperthermophilic archaea, including Pyrococcus and Thermococcus species and Pyrobaculum aerophilum, degrade glucose and glucose polymers by modified versions of the Embden-Meyerhof pathway, whereas thermoacidophilic archaea, e.g., Picrophilus torridus and Sulfolobus species, utilize modified Entner-Doudoroff (ED) pathways, namely, the nonphosphorylative ED pathway and the branched ED pathway, respectively (3,4).…”

Section: Importancementioning

confidence: 99%

Glucose Metabolism and Acetate Switch in Archaea: the Enzymes in Haloferax volcanii

et al. 2021

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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-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. Further, it is shown 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, of anaplerosis, and report 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-CoA and in anaplerosis. Phylogenetic analyses of several enzymes indicate various lateral gene transfer events from bacteria to haloarchaea. Further, 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 on glucose degradation, anaplerosis and acetate switch in H. volcanii and thus expand our understanding of the unusual sugar metabolism in archaea.

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Section: Importancementioning

confidence: 99%

Glucose Metabolism and Acetate Switch in Archaea: the Enzymes in Haloferax volcanii

et al. 2021

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“…The most successful commercial examples of extremophilic products are the DNA manipulating enzymes ( Taq polymerase, Vent ® polymerase), and numerous other extremophilic enzymes that play important roles in commercial drug biosynthesis, household products and large-scale biotransformations (Antranikian and Egorova 2007 ). Extremophilic organisms also continue to yield novel metabolic pathways, enzymes and control elements (e.g., Bräsen et al 2014 ; Weisse et al 2016 ; Saitsev et al 2018 ; Johnsen et al 2019 , 2020 , 2023 ; Stracke et al 2020 ; Kuprat et al 2021 ; Lewis et al 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Extremophiles in a changing world

Cowan,

Albers,

Antranikian

et al. 2024

Extremophiles

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Extremophiles and their products have been a major focus of research interest for over 40 years. Through this period, studies of these organisms have contributed hugely to many aspects of the fundamental and applied sciences, and to wider and more philosophical issues such as the origins of life and astrobiology. Our understanding of the cellular adaptations to extreme conditions (such as acid, temperature, pressure and more), of the mechanisms underpinning the stability of macromolecules, and of the subtleties, complexities and limits of fundamental biochemical processes has been informed by research on extremophiles. Extremophiles have also contributed numerous products and processes to the many fields of biotechnology, from diagnostics to bioremediation. Yet, after 40 years of dedicated research, there remains much to be discovered in this field. Fortunately, extremophiles remain an active and vibrant area of research. In the third decade of the twenty-first century, with decreasing global resources and a steadily increasing human population, the world’s attention has turned with increasing urgency to issues of sustainability. These global concerns were encapsulated and formalized by the United Nations with the adoption of the 2030 Agenda for Sustainable Development and the presentation of the seventeen Sustainable Development Goals (SDGs) in 2015. In the run-up to 2030, we consider the contributions that extremophiles have made, and will in the future make, to the SDGs.

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Sugar alcohol degradation in Archaea: uptake and degradation of mannitol and sorbitol in Haloarcula hispanica

Ortjohann,

Schönheit

2024

Extremophiles

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The halophilic archaeon Haloarcula hispanica utilizes the sugar alcohols mannitol and sorbitol as carbon and energy sources. Genes, enzymes, and transcriptional regulators involved in uptake and degradation of these sugar alcohols were identified by growth experiments with deletion mutants and enzyme characterization. It is shown that both mannitol and sorbitol are taken up via a single ABC transporter of the CUT1 transporter family. Then, mannitol and sorbitol are oxidized to fructose by two distinct dehydrogenases. Fructose is further phosphorylated to fructose-1-phosphate by a haloarchaeal ketohexokinase, providing the first evidence for a physiological function of ketohexokinase in prokaryotes. Finally, fructose-1-phosphate is phosphorylated via fructose-1-phosphate kinase to fructose-1,6-bisphosphate, which is cleaved to triosephosphates by a Class I fructose-1,6-bisphosphate aldolase. Two distinct transcriptional regulators, acting as activators, have been identified: an IclR-like regulator involved in activating genes for sugar alcohol uptake and oxidation to fructose, and a GfcR-like regulator that likely activates genes involved in the degradation of fructose to pyruvate. This is the first comprehensive analysis of a sugar alcohol degradation pathway in Archaea.

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Insights into the Substrate Specificity of Archaeal Entner–Doudoroff Aldolases: The Structures of Picrophilus torridus 2-Keto-3-deoxygluconate Aldolase and Sulfolobus solfataricus 2-Keto-3-deoxy-6-phosphogluconate Aldolase in Complex with 2-Keto-3-deoxy-6-phosphogluconate

Cited by 6 publications

References 25 publications

Glucose Metabolism and Acetate Switch in Archaea: the Enzymes in Haloferax volcanii

Glucose Metabolism and Acetate Switch in Archaea: the Enzymes in Haloferax volcanii

Extremophiles in a changing world

Sugar alcohol degradation in Archaea: uptake and degradation of mannitol and sorbitol in Haloarcula hispanica

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