Predicting the accumulation of storage compounds by Rhodococcus jostii RHA1 in the feast-famine growth cycles using genome-scale flux balance analysis

PLoS One. 2018 Mar 1;13(3):e0191835. doi: 10.1371/journal.pone.0191835. eCollection 2018.

Abstract

Feast-famine cycles in biological wastewater resource recovery systems select for bacterial species that accumulate intracellular storage compounds such as poly-β-hydroxybutyrate (PHB), glycogen, and triacylglycerols (TAG). These species survive better the famine phase and resume rapid substrate uptake at the beginning of the feast phase faster than microorganisms unable to accumulate storage. However, ecophysiological conditions favouring the accumulation of either storage compounds remain to be clarified, and predictive capabilities need to be developed to eventually rationally design reactors producing these compounds. Using a genome-scale metabolic modelling approach, the storage metabolism of Rhodococcus jostii RHA1 was investigated for steady-state feast-famine cycles on glucose and acetate as the sole carbon sources. R. jostii RHA1 is capable of accumulating the three storage compounds (PHB, TAG, and glycogen) simultaneously. According to the experimental observations, when glucose was the substrate, feast phase chemical oxygen demand (COD) accumulation was similar for the three storage compounds; when acetate was the substrate, however, PHB accumulation was 3 times higher than TAG accumulation and essentially no glycogen was accumulated. These results were simulated using the genome-scale metabolic model of R. jostii RHA1 (iMT1174) by means of flux balance analysis (FBA) to determine the objective functions capable of predicting these behaviours. Maximization of the growth rate was set as the main objective function, while minimization of total reaction fluxes and minimization of metabolic adjustment (environmental MOMA) were considered as the sub-objective functions. The environmental MOMA sub-objective performed better than the minimization of total reaction fluxes sub-objective function at predicting the mixture of storage compounds accumulated. Additional experiments with 13C-labelled bicarbonate (HCO3-) found that the fluxes through the central metabolism reactions during the feast phases were similar to the ones during the famine phases on acetate due to similarity in the carbon sources in the feast and famine phases (i.e., acetate and poly-β-hydroxybutyrate, respectively); this suggests that the environmental MOMA sub-objective function could be used to analyze successive environmental conditions such as the feast and famine cycles while the metabolically similar carbon sources are taken up by microorganisms.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Oxygen Demand Analysis
  • Computer Simulation
  • Genome, Bacterial
  • Glucose / metabolism
  • Glycogen / metabolism
  • Hydroxybutyrates / metabolism
  • Metabolic Flux Analysis
  • Metabolic Networks and Pathways
  • Models, Biological
  • Polyesters / metabolism
  • Rhodococcus / genetics
  • Rhodococcus / growth & development*
  • Rhodococcus / metabolism
  • Wastewater / microbiology*

Substances

  • Hydroxybutyrates
  • Polyesters
  • Waste Water
  • poly-beta-hydroxybutyrate
  • Glycogen
  • Glucose

Grants and funding

This work was supported by the Natural Sciences and Engineering Research Council of Canada (NSERC) Discovery Grant awarded to DF (RGPIN 341393-11). MT is grateful to the McGill Engineering Doctoral Award (MEDA) for the financial support of his Ph.D. study. Labelled amino acid measurements were performed at the Rosalind and Morris Goodman Cancer Research Centre Metabolomics Core Facility supported by the Canada Foundation of Innovation, The Dr. John R. and Clara M. Fraser Memorial Trust, the Terry Fox Foundation (TFF Oncometabolism Team Grant 116128), and McGill University. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.