Genome-scale metabolic model driven design of a defined medium for Campylobacter jejuni M1cam
| Autor: | Fauzy Nasher, Noemi Tejera, Lisa Crossman, Dipali Singh, Bruce M. Pearson, John Wain, Bilal Djeghout, Emily Stoakes, Mark G. Poolman |
|---|---|
| Přispěvatelé: | Apollo - University of Cambridge Repository |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Microbiology (medical)
Fastidious organism Auxotrophy lcsh:QR1-502 auxotrophy Metabolic network medicine.disease_cause Campylobacter jejuni Microbiology lcsh:Microbiology 03 medical and health sciences chemistry.chemical_compound metabolic network medicine genome-scale metabolic model Organism 030304 developmental biology Original Research 0303 health sciences Methionine biology 030306 microbiology Campylobacter defined growth media linear programming biology.organism_classification Chemically defined medium chemistry Biochemistry metabolism |
| Zdroj: | Frontiers in Microbiology Frontiers in Microbiology, Vol 11 (2020) |
| Popis: | Campylobacter jejuni, the most frequent cause of food-borne bacterial gastroenteritis, is a fastidious organism when grown in the laboratory. Oxygen is required for growth, despite the presence of the metabolic mechanism for anaerobic respiration. Amino acid auxotrophies are variably reported and energy metabolism can occur through several electron donor/acceptor combinations. Overall, the picture is one of a flexible, but vulnerable metabolism. To understand Campylobacter metabolism, we have constructed a fully curated, metabolic model for the reference organism M1 (our variant is M1cam) and validated it through laboratory experiments. Our results show that M1cam is auxotrophic for methionine, niacinamide, and pantothenate. There are complete biosynthesis pathways for all amino acids except methionine and it can produce energy, but not biomass, in the absence of oxygen. M1cam will grow in DMEM/F-12 defined media but not in the previously published Campylobacter specific defined media tested. Using the model, we identified potential auxotrophies and substrates that may improve growth. With this information, we designed simple defined media containing inorganic salts, the auxotrophic substrates, L-methionine, niacinamide, and pantothenate, pyruvate and additional amino acids L-cysteine, L-serine, and L-glutamine for growth enhancement. Our defined media supports a 1.75-fold higher growth rate than Brucella broth after 48 h at 37°C and sustains the growth of other Campylobacter jejuni strains. This media can be used to design reproducible assays that can help in better understanding the adaptation, stress resistance, and the virulence mechanisms of this pathogen. We have shown that with a well-curated metabolic model it is possible to design a media to grow this fastidious organism. This has implications for the investigation of new Campylobacter species defined through metagenomics, such as C. infans. |
| Databáze: | OpenAIRE |
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