Metabolomic and network analysis of astaxanthin-producing Haematococcus pluvialis under various stress conditions

Various combinations of acetate (Ac), Fe(2+) and high light (HL) stress conditions were evaluated to maximize astaxanthin accumulation and biomass production in Haematococcus pluvialis, and then GC-MS and LC-MS based metabolomics were applied to determine molecular mechanisms responsible for enhancing astaxanthin accumulation under the stress conditions. With the optimized analytical protocols, the GC-MS and LC-MS analyses allowed identification of 93 stable and 24 unstable intracellular metabolites from H. pluvialis, respectively. In addition, a metabolic network was constructed based on GC-MS metabolomic datasets using a weighted correlation network analysis (WGCNA) approach. The network analysis uncovered 2, 1 and 1 distinguished metabolic modules highly associated with HL, Fe(2+) & HL, and Ac & Fe(2+) & HL conditions, respectively. Finally, LC-MS analysis found that AKG, Glu and R5P may be metabolites associated with the Fe(2+) & HL condition. The study provided the first metabolomic view of cell growth and astaxanthin accumulation in H. pluvialis.