Project description:These data belong to a metabolic engineering project that introduces the reductive glycine pathway for formate assimilation in Cupriavidus necator. As part of this project we performed short-term evolution of the bacterium Cupriavidus necator H16 to grow on glycine as sole carbon and energy source. Some mutations in a putiative glycine transporting systems facilitated growth, and we performed transcriptomics on the evolved strain growing on glycine. Analysis of these transcriptomic data lead us to the discovery of a glycine oxidase (DadA6), which we experimentally demonstrated to play a key role in the glycine assimilation pathay in C. necator.
Project description:Previously isolated spontaneous mutants of Rhodobacter sphaeroides, which were initially compromised in their ability to assimilate carbon dioxide through the reductive pentose phosphate pathway, were found to exhibit abnormal nitrogenase gene regulation as well as altered patterns of nitrogenase enzymatic activity. The genomes of these strains were studied by whole genome pyrosequencing and whole genome microarray analysis to identify possible loci responsible for the observed phenotypes. 3 R.sphaeroides strains were grown in matching conditions to reveal strain-specific gene expression
Project description:Previously isolated spontaneous mutants of Rhodobacter sphaeroides, which were initially compromised in their ability to assimilate carbon dioxide through the reductive pentose phosphate pathway, were found to exhibit abnormal nitrogenase gene regulation as well as altered patterns of nitrogenase enzymatic activity. The genomes of these strains were studied by whole genome pyrosequencing and whole genome microarray analysis to identify possible loci responsible for the observed phenotypes.
2013-05-22 | GSE47149 | GEO
Project description:Transcriptome and genome resequencing data of Aurantiochytrium sp. TWZ-97 with Reductive Glycine pathway (rGlyP)
Project description:One-carbon (C1) feedstocks like formate could be energetically efficient substrates for sustainable microbial production of food, fuels and chemicals. Here, we replace the native energy-inefficient Calvin-Benson-Bassham (CBB) cycle in Cupriavidus necator with the more energy-efficient reductive glycine pathway for growth on formate and CO2. In chemostats, our engineered strain reaches a 17% higher biomass yield than the wild type, or any natural formatotroph using the Calvin cycle. This demonstrates the potential of synthetic metabolism to realize sustainable, bio-based production.
2025-02-15 | PXD059545 | Pride
Project description:One-carbon fixation via the reductive glycine pathway exceeds yield of the Calvin cycle in bacteria
Project description:The Wood-Ljungdahl pathway in acetogens converts C1 compounds, such as CO2 and CO, into acetyl-CoA. Similarly, the glycine synthase pathway assimilates C1 compounds into glycine. Partial glycine synthase genes are widely conserved in the Wood-Ljungdahl pathway gene cluster but functional relationship between the pathways in autotrophic condition remains unknown. To comprehend, we assembled Clostridium drakei genome (5.7-Mbp) with intact glycine synthase pathway and constructed a genome-scale metabolic model, iSL836, predicting increased metabolic flux rates of the Wood-Ljungdahl pathway and the glycine synthase-reductase associated reactions under autotrophic conditions. Along with the observation of significant transcriptional activation of genes in the pathways, surprisingly, 13C-labeling experiments and enzyme activity assays confirmed the strain synthesizes glycine and converts into acetyl-phosphate. This study suggests the Wood-Ljungdahl and the glycine synthase-reductase pathways convert CO2 into acetyl-CoA and acetyl-phosphate, respectively. In our knowledge, this is the first report on co-utilization of the pathways under autotrophic growth in acetogen.
