carbohydrate metabolism and carbon fixation in roseobacter denitrificans och114碳水化合物代谢和碳固定roseobacter denitrificans och114.pdfVIP

Carbohydrate Metabolism and Carbon Fixation in Roseobacter denitrificans OCh114 1 2 2 1 Kuo-Hsiang Tang , Xueyang Feng , Yinjie J. Tang , Robert E. Blankenship * 1 Departments of Biology and Chemistry, Washington University in St. Louis, St. Louis, Missouri, United States of America, 2 Department of Energy, Environment and Chemical Engineering, Washington University in St. Louis, St. Louis, Missouri, United States of America 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 biosy