Unraveling the function of the two EntnerDoudoroff branches in the thermoacidophilic Crenarchaeon Sulfolobussolfataricus P2

Kouril, Theresa and Wieloch, Patricia and Reimann, Julia and Wagner, Michaela and Zaparty, Melanie and Albers, Sonja-Verena and Schomburg, Dietmar and Ruoff, Peter and Siebers, Bettina (2013) Unraveling the function of the two EntnerDoudoroff branches in the thermoacidophilic Crenarchaeon Sulfolobussolfataricus P2. FEBS JOURNAL, 280 (4). pp. 1126-1138. ISSN 1742-464X, 1742-4658

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Abstract

Sulfolobussolfataricus P2 is a thermoacidophilic archaeon that metabolizes glucose and galactose via an unusual branched EntnerDoudoroff (ED) pathway, which is characterized by a non-phosphorylative (np) and a semi-phosphorylative (sp) branch. However, so far the physiological significance of the two pathway branches is unknown. In order to address these questions two key enzymes of the branched ED pathway, the class II glycerate kinase (GK) of the np-ED branch and the 2-keto-3-deoxygluconate kinase (KDGK) of the sp-ED branch in S.solfataricus, were investigated. GK was recombinantly purified and characterized with respect to its kinetic properties. Mg2+ dependent Sso-GK (glycerate+ATP2-phosphoglycerate+ADP) showed unusual regulatory properties, i.e. substrate inhibition and cooperativity by d-glycerate and ATP, and a substrate-inhibition model was established fitting closely to the experimental data. Furthermore, deletion of the sp-ED key enzyme KDGK in S.solfataricus PBL2025 resulted in a similar growth phenotype on glucose as substrate compared with the wild-type. In contrast, the mutant showed strongly increased concentrations of np-ED intermediates whereas the hexose and pentose phosphates as well as trehalose were decreased. Together the results indicate (a) that the np-ED pathway is able to compensate for the missing sp-ED branch in glucose catabolism, (b) that in addition to its catabolic function the sp-ED pathway has an additional although not essential role in providing sugar phosphates for anabolism/gluconeogenesis and (c) that GK, with its unusual regulatory properties, seems to play a major role in controlling the flux between the glycolytic np-ED and the glycolytic/gluconeogenetic sp-ED pathway. Database The amino acid sequence data of GK of S.solfataricus P2 is available in the UniProt Protein Database under the accession number Q7LXP1 (http://www.uniprot.org/uniprot/Q7LXP1).

Item Type: Article
Uncontrolled Keywords: ENTNER-DOUDOROFF PATHWAY; ARCHAEON SULFOLOBUS-SOLFATARICUS; BROAD SUBSTRATE-SPECIFICITY; ATP-DEPENDENT HEXOKINASE; GLYCERATE KINASE; KEY-ENZYME; GLUCOSE-DEHYDROGENASE; THERMOTOGA-MARITIMA; THERMOPROTEUS-TENAX; METABOLIC PATHWAYS; carbohydrate metabolism; EntnerDoudoroff pathway; glycerate kinase; metabolomics; thermophiles
Subjects: 500 Science > 570 Life sciences
Divisions: Biology, Preclinical Medicine > Institut für Anatomie > Lehrstuhl für Molekulare und zelluläre Anatomie
Depositing User: Dr. Gernot Deinzer
Date Deposited: 24 Apr 2020 12:25
Last Modified: 24 Apr 2020 12:25
URI: https://pred.uni-regensburg.de/id/eprint/17195

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