Project description:Acetate is a simple carboxylic acid that is synthesized in various microorganisms. Although acetate toxicity and tolerance have been studied in many microorganisms, little is known about the effects of exogenous acetate on the cell growth of acetogenic bacteria. In this study, we report the phenotypic changes that occurred in the acetogenic bacterium Clostridium sp. AWRP as a result of an adaptive laboratory evolution under acetate challenge. When compared with the wild-type strain, the acetate-adapted strain displayed a tolerance to acetate up to 10 g L-1 and higher biomass yields in batch cultures, although the metabolite profiles greatly varied depending on culture conditions. Interestingly, genome sequencing revealed that the adapted strain harbored three point mutations in the genes encoding an electron-bifurcating hydrogenase, which is crucial to its autotrophic growth on CO2 + H2, in addition to one in the dnaK gene. Transcriptome analysis revealed the global change in the gene expression profile of the acetate-adapted strain. Strikingly, most genes involved in CO2-fixing Wood-Ljungdahl pathway and auxiliary pathways for energy conservation (e.g., Rnf complex, Nfn, etc.) were significantly down-regulated. In addition, we observed that a couple of metabolic pathways associated with dissimilation of nucleosides and carbohydrates were significantly up-regulated in the acetate-adapted strain as well as several amino acid biosynthetic pathways, indicating that the strain might increase its fitness by utilizing organic substrates in response to the down-regulation of carbon fixation. Further investigation into the carbon fixation degeneration of the acetate-adapted strain will provide practical implications in CO2 + H2 fermentation using acetogenic bacteria for long-term continuous fermentation. The transcriptome profiles of the wild-type Clostridium sp. AWRP and its acetate-tolerant derivative 46T-a were compared.
Project description:The possibility of establishing a Clostridium-based biorefinery is an attractive and viable alternative, due to the wide metabolic versatility of these microorganisms. The Bioprocesses and Bioprospecting group of the Universidad Nacional de Colombia has obtained the genome of Clostridium sp. IBUN13A, which has shown the ability to produce solvents from various substrates and postulated the need to expand the knowledge of the physiology of the bacteria, so, this study establishes the differences in the transcriptomic profile of the strain when it is cultivated in glycerol respect to glucose after 24 hours of fermentation. For this, a RNA-seq study was carried out, and some of the genes that increased its expression on glycerol were related to the oxidative route of its consumption. The enrichment analysis of Gene Ontology terms showed that the biological phenomena with the highest representation within the differentially expressed genes correspond to oxidation-reduction processes. This first approach to the global transcriptomic study of glycerol fermentation allows the understanding of the metabolism of the microorganism with the purpose of choosing targets for genetic modification.
Project description:This project is a proteomic comparison of Hyphomicrobium sp. MC8b grown with dichloromethane or with methanol. The datasets were obtained using the annotated genome of Hyphomicrobium sp. MC8b.
