Genome-Scale Metabolic Model Driven Design of a Defined Medium for Campylobacter jejuni M1cam
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 en...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2020-06-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2020.01072/full |
| _version_ | 1819036847926411264 |
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| author | Noemi Tejera Lisa Crossman Lisa Crossman Lisa Crossman Bruce Pearson Emily Stoakes Fauzy Nasher Bilal Djeghout Mark Poolman John Wain Dipali Singh |
| author_facet | Noemi Tejera Lisa Crossman Lisa Crossman Lisa Crossman Bruce Pearson Emily Stoakes Fauzy Nasher Bilal Djeghout Mark Poolman John Wain Dipali Singh |
| author_sort | Noemi Tejera |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-21T08:12:02Z |
| format | Article |
| id | doaj.art-1fd07388498b4e1f9b9ada4904ad7aeb |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-21T08:12:02Z |
| publishDate | 2020-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-1fd07388498b4e1f9b9ada4904ad7aeb2022-12-21T19:10:38ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2020-06-011110.3389/fmicb.2020.01072531661Genome-Scale Metabolic Model Driven Design of a Defined Medium for Campylobacter jejuni M1camNoemi Tejera0Lisa Crossman1Lisa Crossman2Lisa Crossman3Bruce Pearson4Emily Stoakes5Fauzy Nasher6Bilal Djeghout7Mark Poolman8John Wain9Dipali Singh10Microbes in Food Chain, Quadram Institute Biosciences, Norwich Research Park, Norwich, United KingdomMicrobes in Food Chain, Quadram Institute Biosciences, Norwich Research Park, Norwich, United KingdomSequenceAnalysis.co.uk, NRP Innovation Centre, Norwich, United KingdomUniversity of East Anglia, Norwich, United KingdomMicrobes in Food Chain, Quadram Institute Biosciences, Norwich Research Park, Norwich, United KingdomDepartment of Veterinary Medicine, University of Cambridge, Cambridge, United KingdomLondon School of Hygiene and Tropical Medicine, University of London, London, United KingdomMicrobes in Food Chain, Quadram Institute Biosciences, Norwich Research Park, Norwich, United KingdomCell Systems Modelling Group, Oxford Brookes University, Oxford, United KingdomMicrobes in Food Chain, Quadram Institute Biosciences, Norwich Research Park, Norwich, United KingdomMicrobes in Food Chain, Quadram Institute Biosciences, Norwich Research Park, Norwich, United KingdomCampylobacter 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.https://www.frontiersin.org/article/10.3389/fmicb.2020.01072/fullCampylobacter jejunigenome-scale metabolic modellinear programmingdefined growth mediametabolismauxotrophy |
| spellingShingle | Noemi Tejera Lisa Crossman Lisa Crossman Lisa Crossman Bruce Pearson Emily Stoakes Fauzy Nasher Bilal Djeghout Mark Poolman John Wain Dipali Singh Genome-Scale Metabolic Model Driven Design of a Defined Medium for Campylobacter jejuni M1cam Frontiers in Microbiology Campylobacter jejuni genome-scale metabolic model linear programming defined growth media metabolism auxotrophy |
| title | Genome-Scale Metabolic Model Driven Design of a Defined Medium for Campylobacter jejuni M1cam |
| title_full | Genome-Scale Metabolic Model Driven Design of a Defined Medium for Campylobacter jejuni M1cam |
| title_fullStr | Genome-Scale Metabolic Model Driven Design of a Defined Medium for Campylobacter jejuni M1cam |
| title_full_unstemmed | Genome-Scale Metabolic Model Driven Design of a Defined Medium for Campylobacter jejuni M1cam |
| title_short | Genome-Scale Metabolic Model Driven Design of a Defined Medium for Campylobacter jejuni M1cam |
| title_sort | genome scale metabolic model driven design of a defined medium for campylobacter jejuni m1cam |
| topic | Campylobacter jejuni genome-scale metabolic model linear programming defined growth media metabolism auxotrophy |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2020.01072/full |
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