Biochemical characterization of human gluconokinase and the proposed metabolic impact of gluconic Acid as determined by constraint based metabolic network analysis
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Biochemical characterization of human gluconokinase and the proposed metabolic impact of gluconic Acid as determined by constraint based metabolic network analysis. / Rohatgi, Neha; Nielsen, Tine Kragh; Bjørn, Sara Petersen; Axelsson, Ivar; Paglia, Giuseppe; Voldborg, Bjørn Gunnar; Palsson, Bernhard O; Rolfsson, Ottar.
In: PloS one, Vol. 9, No. 6, e98760, 04.06.2014.Research output: Contribution to journal › Journal article › Research › peer-review
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T1 - Biochemical characterization of human gluconokinase and the proposed metabolic impact of gluconic Acid as determined by constraint based metabolic network analysis
AU - Rohatgi, Neha
AU - Nielsen, Tine Kragh
AU - Bjørn, Sara Petersen
AU - Axelsson, Ivar
AU - Paglia, Giuseppe
AU - Voldborg, Bjørn Gunnar
AU - Palsson, Bernhard O
AU - Rolfsson, Ottar
PY - 2014/6/4
Y1 - 2014/6/4
N2 - The metabolism of gluconate is well characterized in prokaryotes where it is known to be degraded following phosphorylation by gluconokinase. Less is known of gluconate metabolism in humans. Human gluconokinase activity was recently identified proposing questions about the metabolic role of gluconate in humans. Here we report the recombinant expression, purification and biochemical characterization of isoform I of human gluconokinase alongside substrate specificity and kinetic assays of the enzyme catalyzed reaction. The enzyme, shown to be a dimer, had ATP dependent phosphorylation activity and strict specificity towards gluconate out of 122 substrates tested. In order to evaluate the metabolic impact of gluconate in humans we modeled gluconate metabolism using steady state metabolic network analysis. The results indicate that significant metabolic flux changes in anabolic pathways linked to the hexose monophosphate shunt (HMS) are induced through a small increase in gluconate concentration. We argue that the enzyme takes part in a context specific carbon flux route into the HMS that, in humans, remains incompletely explored. Apart from the biochemical description of human gluconokinase, the results highlight that little is known of the mechanism of gluconate metabolism in humans despite its widespread use in medicine and consumer products.
AB - The metabolism of gluconate is well characterized in prokaryotes where it is known to be degraded following phosphorylation by gluconokinase. Less is known of gluconate metabolism in humans. Human gluconokinase activity was recently identified proposing questions about the metabolic role of gluconate in humans. Here we report the recombinant expression, purification and biochemical characterization of isoform I of human gluconokinase alongside substrate specificity and kinetic assays of the enzyme catalyzed reaction. The enzyme, shown to be a dimer, had ATP dependent phosphorylation activity and strict specificity towards gluconate out of 122 substrates tested. In order to evaluate the metabolic impact of gluconate in humans we modeled gluconate metabolism using steady state metabolic network analysis. The results indicate that significant metabolic flux changes in anabolic pathways linked to the hexose monophosphate shunt (HMS) are induced through a small increase in gluconate concentration. We argue that the enzyme takes part in a context specific carbon flux route into the HMS that, in humans, remains incompletely explored. Apart from the biochemical description of human gluconokinase, the results highlight that little is known of the mechanism of gluconate metabolism in humans despite its widespread use in medicine and consumer products.
U2 - 10.1371/journal.pone.0098760
DO - 10.1371/journal.pone.0098760
M3 - Journal article
C2 - 24896608
VL - 9
JO - PLoS ONE
JF - PLoS ONE
SN - 1932-6203
IS - 6
M1 - e98760
ER -
ID: 118837120