Rapid accumulation of intracellular 2-keto-3-deoxy-6-phosphogluconate in an Entner-Doudoroff aldolase mutant results in bacteriostasis

Lorra K. Fuhrman, Amy Wanken, Kenneth W. Nickerson, Tyrrell Conway

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The accumulation of 2-keto-3-deoxy-6-phosphogluconate, the key intermediate of the Entner-Doudoroff pathway, has long been thought to inhibit growth of bacteria, but careful measurements of 2-keto-3-deoxy-6-phosphogluconate accumulation by growing cells and the correlation of intracellular 2-keto-3-deoxy-6-phosphogluconate levels to growth inhibition had not been made. A system designed for this purposed was developed in Escherichia coli strains, allowing 2-keto-3-deoxy-6-phosphogluconate accumulation to be experimentally induced and measured by extraction of the cell pool. Addition of gluconate to a strain which lacked 2-keto-3-deoxy-6-phosphogluconate aldolase and overproduced 6-phosphogluconate dehydratase resulted in an increase in the intracellular concentration of 2-keto-3-deoxy-6-phosphogluconate from undetectable levels to 2.0 mM within 15 s, as measured by anion-exchange HPLC. The accumulation of 2-keto-3-deoxy-6-phosphogluconate was correlated with an immediate and significant decrease in growth; this inhibition was determined to be bacteriostatic and not bactericidal. It had been proposed that the mechanism of 2-keto-3-deoxy-6-phosphogluconate toxicity involves competitive inhibition of 6-phosphogluconate dehydrogenase and the consequent block of the pentose phosphate pathway. An experiment addressing this hypothesis failed to provide any supporting data.

Original languageEnglish (US)
Pages (from-to)261-266
Number of pages6
JournalFEMS Microbiology Letters
Volume159
Issue number2
DOIs
StatePublished - Feb 15 1998

Fingerprint

Fructose-Bisphosphate Aldolase
phosphogluconate dehydratase
phospho-2-keto-3-deoxy-gluconate aldolase
Growth
Phosphogluconate Dehydrogenase
Pentose Phosphate Pathway
2-keto-3-deoxy-6-phosphogluconate
Anions
High Pressure Liquid Chromatography
Escherichia coli
Bacteria

Keywords

  • 2-Keto-3-deoxy-6-phosphogluconate accumulation
  • Bacteriostasis
  • Entner-Doudoroff pathway

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology
  • Genetics

Cite this

Rapid accumulation of intracellular 2-keto-3-deoxy-6-phosphogluconate in an Entner-Doudoroff aldolase mutant results in bacteriostasis. / Fuhrman, Lorra K.; Wanken, Amy; Nickerson, Kenneth W.; Conway, Tyrrell.

In: FEMS Microbiology Letters, Vol. 159, No. 2, 15.02.1998, p. 261-266.

Research output: Contribution to journalArticle

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abstract = "The accumulation of 2-keto-3-deoxy-6-phosphogluconate, the key intermediate of the Entner-Doudoroff pathway, has long been thought to inhibit growth of bacteria, but careful measurements of 2-keto-3-deoxy-6-phosphogluconate accumulation by growing cells and the correlation of intracellular 2-keto-3-deoxy-6-phosphogluconate levels to growth inhibition had not been made. A system designed for this purposed was developed in Escherichia coli strains, allowing 2-keto-3-deoxy-6-phosphogluconate accumulation to be experimentally induced and measured by extraction of the cell pool. Addition of gluconate to a strain which lacked 2-keto-3-deoxy-6-phosphogluconate aldolase and overproduced 6-phosphogluconate dehydratase resulted in an increase in the intracellular concentration of 2-keto-3-deoxy-6-phosphogluconate from undetectable levels to 2.0 mM within 15 s, as measured by anion-exchange HPLC. The accumulation of 2-keto-3-deoxy-6-phosphogluconate was correlated with an immediate and significant decrease in growth; this inhibition was determined to be bacteriostatic and not bactericidal. It had been proposed that the mechanism of 2-keto-3-deoxy-6-phosphogluconate toxicity involves competitive inhibition of 6-phosphogluconate dehydrogenase and the consequent block of the pentose phosphate pathway. An experiment addressing this hypothesis failed to provide any supporting data.",
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