ABSTRACT
BACKGROUND: The green microalga Dunaliella salina accumulates a high proportion of ß-carotene during abiotic stress conditions. To better understand the intracellular flux distribution leading to carotenoid accumulation, this work aimed at reconstructing a carbon core metabolic network for D. salina CCAP 19/18 based on the recently published nuclear genome and its validation with experimental observations and literature data. RESULTS: The reconstruction resulted in a network model with 221 reactions and 212 metabolites within three compartments: cytosol, chloroplast and mitochondrion. The network was implemented in the MATLAB toolbox CellNetAnalyzer and checked for feasibility. Furthermore, a flux balance analysis was carried out for different light and nutrient uptake rates. The comparison of the experimental knowledge with the model prediction revealed that the results of the stoichiometric network analysis are plausible and in good agreement with the observed behavior. Accordingly, our model provides an excellent tool for investigating the carbon core metabolism of D. salina. CONCLUSIONS: The reconstructed metabolic network of D. salina presented in this work is able to predict the biological behavior under light and nutrient stress and will lead to an improved process understanding for the optimized production of high-value products in microalgae.
Subject(s)
Carbon/metabolism , Chlorophyta/metabolism , Microalgae/metabolism , Carbon/chemistry , Carotenoids/chemistry , Carotenoids/metabolism , Chlorophyta/chemistry , Chlorophyta/radiation effects , Chloroplasts/chemistry , Chloroplasts/metabolism , Cytosol/chemistry , Cytosol/metabolism , Light , Metabolic Networks and Pathways , Microalgae/chemistry , Microalgae/radiation effects , Mitochondria/chemistry , Mitochondria/metabolism , Models, Biological , Stress, PhysiologicalABSTRACT
The primary commercial product from the green microalgae Dunaliella salina is ß-carotene. After extracting the lipophilic fraction containing this red-orange pigment, an algal residue remains. As the carotenogenesis is induced by light stress with simultaneous nitrogen depletion, the protein content is low and the remnant is comprised largely of storage carbohydrates. In this work, we transformed the defatted remnant directly to the platform chemicals, 5-hydroxy methyl furfural (5-HMF) and levulinic acid (LA), without previous purification or any pretreatment. The batch experiments were carried out in an autoclave under biphasic solvent conditions at 453 K for 1 h using acidic ZSM-5 zeolite as a heterogeneous catalyst. Mixtures of methyl isobutyl ketone (MIBK/H2O) or tetrahydrofuran (THF/H2O/NaCl) with water were used to create the biphasic reactor conditions. The biphasic reaction mixtures helped to increase the 5-HMF yield and simultaneously mitigated the formation of insoluble humins. The carbon yields of 5-HMF and of LA in the MIBK/H2O biphasic system without NaCl were 13.9% and 3.7%, respectively. The highest carbon yield of 5-HMF (34.4%) was achieved by adding NaCl to the reaction mixture containing THF/H2O. The experimentally measured partition ratios of 5-HMF between the two liquid phases were compared to the predictions calculated by the computational method COSMO-RS, which is a quantum chemistry-based method to predict the thermodynamic equilibria of liquid mixtures and the solubilities. The COSMO-RS predicted partition ratios of 5-HMF were in line with the experimentally measured ones.