From Feasting to Fasting: The Arginine Pathway as a Metabolic Switch in Nitrogen-Deprived Chlamydomonas reinhardtii.

The metabolism of the model microalgae Chlamydomonas reinhardtii under nitrogen deprivation is of special interest due to its resulting increment of triacylglycerols (TAGs), that can be applied in biotechnological applications. However, this same condition impairs cell growth, which may limit the microalgae's large applications. Several studies have identified significant physiological and molecular changes that occur during the transition from an abundant to a low or absent nitrogen supply, explaining in detail the differences in the proteome, metabolome and transcriptome of the cells that may be responsible for and responsive to this condition. However, there are still some intriguing questions that reside in the core of the regulation of these cellular responses that make this process even more interesting and complex. In this scenario, we reviewed the main metabolic pathways that are involved in the response, mining and exploring, through a reanalysis of omics data from previously published datasets, the commonalities among the responses and unraveling unexplained or non-explored mechanisms of the possible regulatory aspects of the response. Proteomics, metabolomics and transcriptomics data were reanalysed using a common strategy, and an in silico gene promoter motif analysis was performed. Together, these results identified and suggested a strong association between the metabolism of amino acids, especially arginine, glutamate and ornithine pathways to the production of TAGs, via the de novo synthesis of lipids. Furthermore, our analysis and data mining indicate that signalling cascades orchestrated with the indirect participation of phosphorylation, nitrosylation and peroxidation events may be essential to the process. The amino acid pathways and the amount of arginine and ornithine available in the cells, at least transiently during nitrogen deprivation, may be in the core of the post-transcriptional, metabolic regulation of this complex phenomenon. Their further exploration is important to the discovery of novel advances in the understanding of microalgae lipids' production.

A Multi-Omics Extraction Method for the In-Depth Analysis of Synchronized Cultures of the Green Alga Chlamydomonas reinhardtii.

Microalgae have been the focus of research for their applications in the production of high value compounds, food and fuel. Moreover, they are valuable photosynthetic models facilitating the understanding of the basic cellular processes. System wide studies enable comprehensive and in-depth understanding of molecular functions of the organisms. However, multiple independent samples and protocols are required for proteomics, lipidomics and metabolomics studies introducing higher error and variability. A robust high throughput extraction method for the simultaneous extraction of chlorophyll, lipids, metabolites, proteins and starch from a single sample of the green alga Chlamydomonas reinhardtii is presented here. The illustrated experimental setup is for Chlamydomonas cultures synchronized using 12 h/12 h light/dark conditions. Samples were collected over a 24 h cell cycle to demonstrate that the metabolites, lipids and starch data obtained using various analytical platforms are well conformed. Furthermore, protein samples collected using the same extraction protocol were used to conduct detailed proteomics analysis to evaluate their quality and reproducibility. Based on the data, it can be inferred that the illustrated method provides a robust and reproducible approach to advance understanding of various biochemical pathways and their functions with greater confidence for both basic and applied research.

Anaerobic phototrophic processes of hydrogen production by different strains of microalgae Chlamydomonas sp.

Hydrogen is an abundant element and a non-polluting fuel that can be biologically produced by microalgae. The aim of this research was to investigate biological hydrogen production by Chlamydomonas reinhardtii (CC425) and Chlamydomonas moewusii (SAG 24.91) by direct biophotolysis in batch cultures. Strains were cultivated in TAP growth medium (pH 7.2) in two phases: in the first stage, cultures were maintained in an aerobic condition until the middle of the exponential phase; in the second stage, the biomass was transferred to closed anaerobic photobioreactors under sulfur deprived. Gas chromatography and Gompertz model were used to measure the hydrogen production and hydrogen production rate, respectively. We noticed that maximum hydrogen production by biomass of C. reinhardtii was 5.95 ± 0.88 µmol mg-1 and the productivity was 17.02 ± 3.83 µmol L-1 h-1, with hydrogen production five times higher than C. moewusii, approximately, though, C. moewusii obtained a higher ethanol yield compared to C. reinhardtii. The hydrogen production method, with the cultivation of strains in two different phases and sulfur deprivation, was effective for obtaining of biohydrogen for Chlamydomonas; however, it depends on the species, strain and growth conditions.