The phosphoenolpyruvate carboxykinase also catalyzes C3 carboxylation at the interface of glycolysis and the TCA cycle of Bacillus subtilis

Nicola Zamboni, Hannu Maaheimo, Thomas Szyperski, Hans-Peter Hohmann, Uwe Sauer (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

39 Citations (Scopus)

Abstract

Quantitative physiological characterization and isotopic tracer experiments revealed that pyruvate kinase mutants of Bacillus subtilis produced significantly more CO2 from glucose in the tricarboxylic acid cycle than is explained by the remaining conversion of phosphoenolpyruvate (PEP) to pyruvate catalyzed by the phosphotransferase system. We show here that this additional catabolic flux into the tricarboxylic acid cycle was catalyzed by the PEP carboxykinase. In contrast to its normal role in gluconeogenesis, PEP carboxykinase can operate in the reverse direction from PEP to oxaloacetate upon knockout of pyruvate kinase in a riboflavin-producing B. subtilis strain and in wild-type 168. At least in the industrial strain, we demonstrate the additional capacity of PEP carboxykinase to function as a substitute anaplerotic reaction when the normal pyruvate carboxylase is inactivated. Presumably as a consequence of the unfavorable kinetics of an ATP-synthesizing anaplerotic PEP carboxykinase reaction, such pyruvate carboxylase mutants grow slowly or, as in the case of wild-type 168, not at all.

Original languageEnglish
Pages (from-to)277-284
Number of pages8
JournalMetabolic Engineering
Volume6
Issue number4
DOIs
Publication statusPublished - 2004
MoE publication typeA1 Journal article-refereed

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Carboxylation
Phosphoenolpyruvate
Glycolysis
Bacilli
Bacillus subtilis
Enzyme kinetics
Adenosinetriphosphate
Pyruvate Carboxylase
Glucose
Pyruvate Kinase
Citric Acid Cycle
Fluxes
Oxaloacetic Acid
Gluconeogenesis
Riboflavin
Experiments
Pyruvic Acid
Phosphotransferases
Adenosine Triphosphate
Kinetics

Keywords

  • metabolic networks
  • metabolism
  • catalysis
  • catalysts
  • tricarboxylic acid cycle

Cite this

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title = "The phosphoenolpyruvate carboxykinase also catalyzes C3 carboxylation at the interface of glycolysis and the TCA cycle of Bacillus subtilis",
abstract = "Quantitative physiological characterization and isotopic tracer experiments revealed that pyruvate kinase mutants of Bacillus subtilis produced significantly more CO2 from glucose in the tricarboxylic acid cycle than is explained by the remaining conversion of phosphoenolpyruvate (PEP) to pyruvate catalyzed by the phosphotransferase system. We show here that this additional catabolic flux into the tricarboxylic acid cycle was catalyzed by the PEP carboxykinase. In contrast to its normal role in gluconeogenesis, PEP carboxykinase can operate in the reverse direction from PEP to oxaloacetate upon knockout of pyruvate kinase in a riboflavin-producing B. subtilis strain and in wild-type 168. At least in the industrial strain, we demonstrate the additional capacity of PEP carboxykinase to function as a substitute anaplerotic reaction when the normal pyruvate carboxylase is inactivated. Presumably as a consequence of the unfavorable kinetics of an ATP-synthesizing anaplerotic PEP carboxykinase reaction, such pyruvate carboxylase mutants grow slowly or, as in the case of wild-type 168, not at all.",
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The phosphoenolpyruvate carboxykinase also catalyzes C3 carboxylation at the interface of glycolysis and the TCA cycle of Bacillus subtilis. / Zamboni, Nicola; Maaheimo, Hannu; Szyperski, Thomas; Hohmann, Hans-Peter; Sauer, Uwe (Corresponding Author).

In: Metabolic Engineering, Vol. 6, No. 4, 2004, p. 277-284.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Maaheimo, Hannu

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AU - Hohmann, Hans-Peter

AU - Sauer, Uwe

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AB - Quantitative physiological characterization and isotopic tracer experiments revealed that pyruvate kinase mutants of Bacillus subtilis produced significantly more CO2 from glucose in the tricarboxylic acid cycle than is explained by the remaining conversion of phosphoenolpyruvate (PEP) to pyruvate catalyzed by the phosphotransferase system. We show here that this additional catabolic flux into the tricarboxylic acid cycle was catalyzed by the PEP carboxykinase. In contrast to its normal role in gluconeogenesis, PEP carboxykinase can operate in the reverse direction from PEP to oxaloacetate upon knockout of pyruvate kinase in a riboflavin-producing B. subtilis strain and in wild-type 168. At least in the industrial strain, we demonstrate the additional capacity of PEP carboxykinase to function as a substitute anaplerotic reaction when the normal pyruvate carboxylase is inactivated. Presumably as a consequence of the unfavorable kinetics of an ATP-synthesizing anaplerotic PEP carboxykinase reaction, such pyruvate carboxylase mutants grow slowly or, as in the case of wild-type 168, not at all.

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KW - catalysis

KW - catalysts

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