Abstract
Introduction
The use of urea as a nitrogen (N) source by Chlorophytes usually enhances biomass and lipid production when compared to ammonium (NH4+). However, the metabolic shifts displayed by Chlamydomonas reinhardtii growing with this organic N source are not known.
Objectives
This study aimed: (i) to characterize the metabolism of C. reinhardtii cultivated in media containing only urea as N source as well as combined with different NH4+ ratios; (ii) to understand how metabolism respond to urea availability.
Methods
Specific quantification of metabolites using 96-well microplates, and high-performance liquid chromatography combined with non-targeted metabolite profiling by gas chromatography (GC)–time-of-flight (TOF)-mass spectrometry (MS) were used in this study. In addition, GC analysis was used to determine fatty acid profiling.
Results
The use of urea did not alter the growth rate in comparison with NH4+. Interestingly, the cell number decreased and the cell size increased proportionally with urea availability. Furthermore, chlorophyll, protein and lipid contents increased with the amount of urea. Regarding the fatty acid profile, oleic acid (C18:1 w8) decreased with amount of urea, while linoleic acid (C18:2 w6) doubled in urea-containing medium.
Conclusions
These results indicate that urea promotes remarkable adjustments in metabolism, without drastic changes in biomass, promoting changes in carbohydrate and amino acid metabolism, as well as in lipids production and fatty acid profile.
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Acknowledgements
This work was supported by Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) [Grant APQ-01357-14, APQ-01671-15 and RED-00053-16] and Max Planck Society to WLA. Research fellowships granted by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) to ANN, WLA, MM, and ASM and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) to ADB and RMR are also gratefully acknowledged. M.G.M.V.V. was supported by scholarships from CAPES/Fundação de Amparo à Pesquisa do estado de Minas Gerais (FAPEMIG) (BPD-00514-14) and CAPES (PNPD-1638006). The authors wish to thank the NUBIOMOL-UFV for providing the facilities for the analysis of this work.
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ADB and ANN conceived and designed research. ADB, RMR, ASM, BAS and PFG conducted experiments. ADB, MM and MGMVV analyzed data. ADB and ANN wrote the manuscript. LC and WLA revised the manuscript. All authors read and approved the final manuscript.
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Batista, A.D., Rosa, R.M., Machado, M. et al. Increased urea availability promotes adjustments in C/N metabolism and lipid content without impacting growth in Chlamydomonas reinhardtii. Metabolomics 15, 31 (2019). https://doi.org/10.1007/s11306-019-1496-3
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DOI: https://doi.org/10.1007/s11306-019-1496-3