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Combined fluxomics and transcriptomics analysis of glucose catabolism via a partially cyclic pentose phosphate pathway in Gluconobacter oxydans 621H.
Appl Environ Microbiol. 2013 Apr; 79(7):2336-48.AE

Abstract

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.

Authors+Show Affiliations

Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich, Jülich, Germany.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

23377928

Citation

Hanke, Tanja, et al. "Combined Fluxomics and Transcriptomics Analysis of Glucose Catabolism Via a Partially Cyclic Pentose Phosphate Pathway in Gluconobacter Oxydans 621H." Applied and Environmental Microbiology, vol. 79, no. 7, 2013, pp. 2336-48.
Hanke T, Nöh K, Noack S, et al. Combined fluxomics and transcriptomics analysis of glucose catabolism via a partially cyclic pentose phosphate pathway in Gluconobacter oxydans 621H. Appl Environ Microbiol. 2013;79(7):2336-48.
Hanke, T., Nöh, K., Noack, S., Polen, T., Bringer, S., Sahm, H., Wiechert, W., & Bott, M. (2013). Combined fluxomics and transcriptomics analysis of glucose catabolism via a partially cyclic pentose phosphate pathway in Gluconobacter oxydans 621H. Applied and Environmental Microbiology, 79(7), 2336-48. https://doi.org/10.1128/AEM.03414-12
Hanke T, et al. Combined Fluxomics and Transcriptomics Analysis of Glucose Catabolism Via a Partially Cyclic Pentose Phosphate Pathway in Gluconobacter Oxydans 621H. Appl Environ Microbiol. 2013;79(7):2336-48. PubMed PMID: 23377928.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Combined fluxomics and transcriptomics analysis of glucose catabolism via a partially cyclic pentose phosphate pathway in Gluconobacter oxydans 621H. AU - Hanke,Tanja, AU - Nöh,Katharina, AU - Noack,Stephan, AU - Polen,Tino, AU - Bringer,Stephanie, AU - Sahm,Hermann, AU - Wiechert,Wolfgang, AU - Bott,Michael, Y1 - 2013/02/01/ PY - 2013/2/5/entrez PY - 2013/2/5/pubmed PY - 2013/8/29/medline SP - 2336 EP - 48 JF - Applied and environmental microbiology JO - Appl. Environ. Microbiol. VL - 79 IS - 7 N2 - 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. SN - 1098-5336 UR - https://www.unboundmedicine.com/medline/citation/23377928/Combined_fluxomics_and_transcriptomics_analysis_of_glucose_catabolism_via_a_partially_cyclic_pentose_phosphate_pathway_in_Gluconobacter_oxydans_621H_ L2 - http://aem.asm.org/cgi/pmidlookup?view=long&pmid=23377928 DB - PRIME DP - Unbound Medicine ER -