This dataset presents the fatty acid composition and biodiesel profile of microalgal biomass cultivated under varying light intensities and CO₂ concentrations. The analysis was performed to investigate the impact of these environmental factors on the quality and composition of biodiesel derived from the biomass. The dataset includes: Fatty acid methyl ester (FAME) profiles of the biomass. Biodiesel property data derived from the FAME profiles. Experimental conditions (light intensities and CO₂ concentrations). Results of statistical analysis comparing fatty acid composition and biodiesel properties across experimental conditions. Analysis Methods:Statistical comparisons were conducted using one-way ANOVA followed by Tukey's post hoc test to identify significant differences among the experimental groups. Microsoft Excel’s built-in tools were utilized to perform the analysis. Purpose:The findings contribute to optimizing cultivation parameters for producing high-quality biodiesel from microalgae, supporting sustainable biofuel production efforts.