The Mass Spectrometry Analysis of Chlamydomonas Reinhardtii During Nitrogen Starvation
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Chlamydomonas reinhardtii is a soil microalgae that has been found to produce useful biopharmaceutical and biofuel material. It is known that mutants of Chlamydomonas reinhardtii produce an abundance of lipid compounds during nitrogen starvation. This project aims to identify possible sources of energy for cellular processes and use mass spectrometry and carbon thirteen isotope labeling to characterize the process of lipid production during nitrogen starvation. To conduct this research, the sta6 mutant of Chlamydomonas reinhardtii was first grown in unlabeled acetate for 120 hours followed by nitrogen starvation in [1-13C] labeled acetate for 72 hours. Mass spectrometry analysis of algal headspace at 24, 48, and 72 hours post-starvation shows that Chlamydomonas reinhardtii utilizes the labeled acetate to obtain carbons from outside the cell and incorporate them into fatty acid tails. Moreover, mass spectrometry characterization of post-starved algal contents revealed unique isotopic distributions for observed diacylglycerols and triacylglycerols. Comparisons of results from mass spectrometry analysis with published transcriptome and pathway analyses allows the identification of key enzymes involved in driving the lipid production during nitrogen starvation.