title: The use of genome-scale metabolic network reconstruction to predict fluxes and equilibrium composition of N-fixing versus C-fixing cells in a diazotrophic cyanobacterium, Trichodesmium erythraeum.
abstract: Computational, genome based predictions of organism phenotypes has enhanced the ability to investigate the biological phenomena that help organisms survive and respond to their environments. In this study, we have created the first genome-scale metabolic network reconstruction of the nitrogen fixing cyanobacterium T. erythraeum and used genome-scale modeling approaches to investigate carbon and nitrogen fluxes as well as growth and equilibrium population composition.
  
  
  
  We created a genome-scale reconstruction of T. erythraeum with 971 reactions, 986 metabolites, and 647 unique genes. We then used data from previous studies as well as our own laboratory data to establish a biomass equation and two distinct submodels that correspond to the two cell types formed by T. erythraeum. We then use flux balance analysis and flux variability analysis to generate predictions for how metabolism is distributed to account for the unique productivity of T. erythraeum. Finally, we used in situ data to constrain the model, infer time dependent population compositions and metabolite production using dynamic Flux Balance Analysis. We find that our model predicts equilibrium compositions similar to laboratory measurements, approximately 15.5% diazotrophs for our model versus 10-20% diazotrophs reported in literature. We also found that equilibrium was the most efficient mode of growth and that equilibrium was stoichiometrically mediated. Moreover, the model predicts that nitrogen leakage is an essential condition of optimality for T. erythraeum; cells leak approximately 29.4% total fixed nitrogen when growing at the optimal growth rate, which agrees with values observed in situ.
  
  
  
  The genome-metabolic network reconstruction allows us to use constraints based modeling approaches to predict growth and optimal cellular composition in T. erythraeum colonies. Our predictions match both in situ and laboratory data, indicating that stoichiometry of metabolic reactions plays a large role in the differentiation and composition of different cell types. In order to realize the full potential of the model, advance modeling techniques which account for interactions between colonies, the environment and surrounding species need to be developed.
authors.affiliation: Department of Chemical and Biological Engineering, Colorado School of Mines, Golden, CO, 80401, USA.
authors.forename: Joseph J
authors.initials: JJ
authors.lastname: Gardner
authors.affiliation: Department of Chemical and Biological Engineering, Colorado School of Mines, Golden, CO, 80401, USA. nboyle@mines.edu.
authors.forename: Nanette R
authors.initials: NR
authors.lastname: Boyle
mesh_terms: D018533:Biomass; D057486:Carbon Cycle; D023281:Genomics / Q000379:methods*; D064688:Metabolic Flux Analysis; D053858:Metabolic Networks and Pathways; D009586:Nitrogen Fixation; D000072359:Trichodesmium / Q000166:cytology / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*
keywords: Constraints based modeling; Diazotrophy; Flux balance analysis; Flux variability analysis; Marine cyanobacteria; Nitrogen cycle