| Summary: | The objective of this study, for the first time, was to optimize Amazonian cyanobacterial culture conditions for improving cell productivity and lipid content, by analyzing the effect of light intensity and nitrogen concentration, for empirically evaluating biodiesel quality parameters. The strains <i>Synechocystis</i> sp. CACIAM05, <i>Microcystis aeruginosa</i> CACIAM08, <i>Pantanalinema rosaneae</i> CACIAM18, and <i>Limnothrix</i> sp. CACIAM25, were previously identified by morphological and molecular analysis (16S rRNA) and were selected based on their production of chlorophyll a and dry cell weight. Then, factorial planning (2<sup>2</sup>) with central points was applied, with light intensity and NaNO<sub>3</sub> concentration as independent variables. As response variables, cell productivity and lipid content were determined. Statistical analysis indicated that for all strains, the independent variables were statistically significant for cell productivity. Analysis of the fatty acid composition demonstrated diversity in the composition of the fatty acid profile from the experimental planning assays of each strain. The Biodiesel Analyzer software predicted the biodiesel quality parameters. CACIAM05 and CACIAM25 obtained better parameters with low levels of light intensity and NaNO<sub>3</sub> concentration, whereas CACIAM08 and CACIAM18 obtained better parameters with low NaNO<sub>3</sub> concentrations and high luminous intensity.
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