Role of the pentose phosphate pathway and the Entner–Doudoroff pathway in glucose metabolism of Gluconobacter oxydans 621H

Richhardt, Janine; Bringer, Stephanie; Bott, Michael

doi:10.1007/s00253-013-4707-2

Cited by 36 publications

(48 citation statements)

References 14 publications

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“…The major route for 6-phosphogluconate metabolism was the PPP, 62% and 93% in growth phases I and II, respectively. This preference of the PPP was also observed by us in a recent analysis of ⌬gnd and ⌬edd-eda mutants of G. oxydans during growth on mannitol (15) or on glucose (43). The prevalence of the PPP as the major catabolic pathway is a feature not often observed in bacteria.…”

Section: Discussionmentioning

confidence: 50%

Combined Fluxomics and Transcriptomics Analysis of Glucose Catabolism via a Partially Cyclic Pentose Phosphate Pathway in Gluconobacter oxydans 621H

Hanke¹,

Nöh²,

Noack³

et al. 2013

Appl Environ Microbiol

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In this study, the distribution and regulation of periplasmic and cytoplasmic carbon fluxes in Gluconobacter oxydans 621H with glucose were studied by 13 C-based metabolic flux analysis ( 13 C-MFA) in combination with transcriptomics and enzyme assays. For 13 C-MFA, cells were cultivated with specifically 13 C-labeled glucose, and intracellular metabolites were analyzed for their labeling pattern by liquid chromatography-mass spectrometry (LC-MS). In growth phase I, 90% of the glucose was oxidized periplasmically to gluconate and partially further oxidized to 2-ketogluconate. Of the glucose taken up by the cells, 9% was phosphorylated to glucose 6-phosphate, whereas 91% was oxidized by cytoplasmic glucose dehydrogenase to gluconate. Additional gluconate was taken up into the cells by transport. Of the cytoplasmic gluconate, 70% was oxidized to 5-ketogluconate and 30% was phosphorylated to 6-phosphogluconate. In growth phase II, 87% of gluconate was oxidized to 2-ketogluconate in the periplasm and 13% was taken up by the cells and almost completely converted to 6-phosphogluconate. Since G. oxydans lacks phosphofructokinase, glucose 6-phosphate can be metabolized only via the oxidative pentose phosphate pathway (PPP) or the Entner-Doudoroff pathway (EDP).13 C-MFA showed that 6-phosphogluconate is catabolized primarily via the oxidative PPP in both phases I and II (62% and 93%) and demonstrated a cyclic carbon flux through the oxidative PPP. The transcriptome comparison revealed an increased expression of PPP genes in growth phase II, which was supported by enzyme activity measurements and correlated with the increased PPP flux in phase II. Moreover, genes possibly related to a general stress response displayed increased expression in growth phase II.

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Section: Discussionmentioning

confidence: 50%

Combined Fluxomics and Transcriptomics Analysis of Glucose Catabolism via a Partially Cyclic Pentose Phosphate Pathway in Gluconobacter oxydans 621H

Hanke¹,

Nöh²,

Noack³

et al. 2013

Appl Environ Microbiol

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“…The exact effects are strongly dependent on the organism, the genetic background of the parent strain, and the environmental growth conditions [185,202,204,[218][219][220]. However, in general, G6PDH-deficient and 6PGDH-deficient prokaryotes exhibit remarkably mild phenotypes.…”

Section: Introductionmentioning

confidence: 99%

Thermococcus kodakarensis : the key to affordable biohydrogen production

Spaans¹

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“…The bacterial strains and plasmids used in this study are listed in Table 1. Escherichia coli strains were cultivated in lysogeny broth (LB) medium (22,23) 4 . The initial pH value of the medium was 6.0.…”

Section: Methodsmentioning

confidence: 99%

“…G. oxydans IFO3293 and G. oxydans 621H cells cultivated with glucose as the carbon source in a bioreactor reach a cell yield of 0.09 g of cells (dry weight) [g cdw ]/g of glucose (4,17). In comparison to these values, E. coli reaches a value of 0.49 g cdw /g of glucose, and Bacillus subtilis reaches a yield of 0.32 g cdw /g of glucose (18,19).…”

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confidence: 95%

“…Important applications are in the production of vitamin C and of 6-amino-L-sorbose, an intermediate in the synthesis of the antidiabetes drug miglitol (3). Only small fractions of the carbon and energy sources are metabolized in the cytoplasm, either via the pentose phosphate pathway or via the Entner-Doudoroff pathway, and it was demonstrated that the former one is of major importance for growth on mannitol and glucose (4,5). Moreover, it was recently shown that the pentose phosphate pathway has partially cyclic operation during growth on glucose (6).…”

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Evidence for a Key Role of Cytochrome bo3 Oxidase in Respiratory Energy Metabolism of Gluconobacter oxydans

Richhardt

Luchterhand

Bringer

et al. 2013

Journal of Bacteriology

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The obligatory aerobic acetic acid bacterium Gluconobacter oxydans oxidizes a variety of substrates in the periplasm by membrane-bound dehydrogenases, which transfer the reducing equivalents to ubiquinone. Two quinol oxidases, cytochrome bo 3 and cytochrome bd, then catalyze transfer of the electrons from ubiquinol to molecular oxygen. In this study, mutants lacking either of these terminal oxidases were characterized. Deletion of the cydAB genes for cytochrome bd had no obvious influence on growth, whereas the lack of the cyoBACD genes for cytochrome bo 3 severely reduced the growth rate and the cell yield. Using a respiration activity monitoring system and adjusting different levels of oxygen availability, hints of a low-oxygen affinity of cytochrome bd oxidase were obtained, which were supported by measurements of oxygen consumption in a respirometer. The H ؉ /O ratio of the ⌬cyoBACD mutant with mannitol as the substrate was 0.56 ؎ 0.11 and more than 50% lower than that of the reference strain (1.26 ؎ 0.06) and the ⌬cydAB mutant (1.31 ؎ 0.16), indicating that cytochrome bo 3 oxidase is the main component for proton extrusion via the respiratory chain. Plasmid-based overexpression of cyoBACD led to increased growth rates and growth yields, both in the wild type and the ⌬cyoBACD mutant, suggesting that cytochrome bo 3 might be a rate-limiting factor of the respiratory chain.

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Role of the pentose phosphate pathway and the Entner–Doudoroff pathway in glucose metabolism of Gluconobacter oxydans 621H

Cited by 36 publications

References 14 publications

Combined Fluxomics and Transcriptomics Analysis of Glucose Catabolism via a Partially Cyclic Pentose Phosphate Pathway in Gluconobacter oxydans 621H

Combined Fluxomics and Transcriptomics Analysis of Glucose Catabolism via a Partially Cyclic Pentose Phosphate Pathway in Gluconobacter oxydans 621H

Thermococcus kodakarensis : the key to affordable biohydrogen production

Evidence for a Key Role of Cytochrome bo3 Oxidase in Respiratory Energy Metabolism of Gluconobacter oxydans

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