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Title: Carbohydrate Metabolism and Carbon Fixation in Roseobacter denitrificans OCh114

Abstract

The Roseobacter clade of aerobic marine proteobacteria, which compose 10–25% of the total marine bacterial community, has been reported to fix CO2, although it has not been determined what pathway is involved. In this study, we report the first metabolic studies on carbohydrate utilization, CO2 assimilation, and amino acid biosynthesis in the phototrophic Roseobacter clade bacterium Roseobacter denitrificans OCh114. We develop a new minimal medium containing defined carbon source(s), in which the requirements of yeast extract reported previously for the growth of R. denitrificans can be replaced by vitamin B12 (cyanocobalamin). Tracer experiments were carried out in R. denitrificans grown in a newly developed minimal medium containing isotopically labeled pyruvate, glucose or bicarbonate as a single carbon source or in combination. Through measurements of 13C-isotopomer labeling patterns in protein-derived amino acids, gene expression profiles, and enzymatic activity assays, we report that: (1) R. denitrificans uses the anaplerotic pathways mainly via the malic enzyme to fix 10–15% of protein carbon from CO2; (2) R. denitrificans employs the Entner-Doudoroff (ED) pathway for carbohydrate metabolism and the non-oxidative pentose phosphate pathway for the biosynthesis of histidine, ATP, and coenzymes; (3) the Embden-Meyerhof-Parnas (EMP, glycolysis) pathway is not active and the enzymatic activity ofmore » 6-phosphofructokinase (PFK) cannot be detected in R. denitrificans; and (4) isoleucine can be synthesized from both threonine-dependent (20% total flux) and citramalate-dependent (80% total flux) pathways using pyruvate as the sole carbon source.« less

Authors:
 [1];  [1];  [1];  [1]
  1. Washington Univ., St. Louis, MO (United States)
Publication Date:
Research Org.:
Washington Univ., St. Louis, MO (United States)
Sponsoring Org.:
USDOE Office of Science (SC); National Aeronautics and Space Administration (NASA)
OSTI Identifier:
1904750
Grant/Contract Number:  
FG02-08ER64694
Resource Type:
Accepted Manuscript
Journal Name:
PLoS ONE
Additional Journal Information:
Journal Volume: 4; Journal Issue: 10; Journal ID: ISSN 1932-6203
Publisher:
Public Library of Science
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Science & Technology - Other Topics; Pyruvate; Carbon dioxide; Isoleucine; Glucose; Biosynthesis; Cobalamins; Threonine; Yeast

Citation Formats

Tang, Kuo-Hsiang, Feng, Xueyang, Tang, Yinjie J., and Blankenship, Robert E. Carbohydrate Metabolism and Carbon Fixation in Roseobacter denitrificans OCh114. United States: N. p., 2009. Web. doi:10.1371/journal.pone.0007233.
Tang, Kuo-Hsiang, Feng, Xueyang, Tang, Yinjie J., & Blankenship, Robert E. Carbohydrate Metabolism and Carbon Fixation in Roseobacter denitrificans OCh114. United States. https://doi.org/10.1371/journal.pone.0007233
Tang, Kuo-Hsiang, Feng, Xueyang, Tang, Yinjie J., and Blankenship, Robert E. Thu . "Carbohydrate Metabolism and Carbon Fixation in Roseobacter denitrificans OCh114". United States. https://doi.org/10.1371/journal.pone.0007233. https://www.osti.gov/servlets/purl/1904750.
@article{osti_1904750,
title = {Carbohydrate Metabolism and Carbon Fixation in Roseobacter denitrificans OCh114},
author = {Tang, Kuo-Hsiang and Feng, Xueyang and Tang, Yinjie J. and Blankenship, Robert E.},
abstractNote = {The Roseobacter clade of aerobic marine proteobacteria, which compose 10–25% of the total marine bacterial community, has been reported to fix CO2, although it has not been determined what pathway is involved. In this study, we report the first metabolic studies on carbohydrate utilization, CO2 assimilation, and amino acid biosynthesis in the phototrophic Roseobacter clade bacterium Roseobacter denitrificans OCh114. We develop a new minimal medium containing defined carbon source(s), in which the requirements of yeast extract reported previously for the growth of R. denitrificans can be replaced by vitamin B12 (cyanocobalamin). Tracer experiments were carried out in R. denitrificans grown in a newly developed minimal medium containing isotopically labeled pyruvate, glucose or bicarbonate as a single carbon source or in combination. Through measurements of 13C-isotopomer labeling patterns in protein-derived amino acids, gene expression profiles, and enzymatic activity assays, we report that: (1) R. denitrificans uses the anaplerotic pathways mainly via the malic enzyme to fix 10–15% of protein carbon from CO2; (2) R. denitrificans employs the Entner-Doudoroff (ED) pathway for carbohydrate metabolism and the non-oxidative pentose phosphate pathway for the biosynthesis of histidine, ATP, and coenzymes; (3) the Embden-Meyerhof-Parnas (EMP, glycolysis) pathway is not active and the enzymatic activity of 6-phosphofructokinase (PFK) cannot be detected in R. denitrificans; and (4) isoleucine can be synthesized from both threonine-dependent (20% total flux) and citramalate-dependent (80% total flux) pathways using pyruvate as the sole carbon source.},
doi = {10.1371/journal.pone.0007233},
journal = {PLoS ONE},
number = 10,
volume = 4,
place = {United States},
year = {Thu Oct 01 00:00:00 EDT 2009},
month = {Thu Oct 01 00:00:00 EDT 2009}
}

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