Technical Abstract: Kinetic analysis was investigated for a carbon-limited medium, with a carbon-to-nitrogen ratio of 5.0, supporting the growth of the 5,8,11,14,17-eicosapentaenoic acid (20:5; omega-3)accumulating fungal organism Pythium irregulare. The productivity and yield parameters at three temperatures, 14, 21, and 28 degrees Celsius, demonstrated growth-coupled synthesis for lipid-free biomass growth and lipid accumulation. For this system, the maximum specific growth rate and theoretical maximum biomass yield based on logistic growth kinetics were used to determine an activation energy of the growth process, of 36.5 kilojoules per mol. At 14, 21, and 28 degrees Celsius, peak lipid yield occurred after culturing for 7, 4, and 3 days, respectively, with peak lipid yields of 8.14, 12.8, and 6.69 grams lipid per 100 grams of glucose. At these peak yields, the maximum lipid-free biomass productivity was achieved at the colder 14 degrees Celsius temperature as well as an increased concentration of eicosapentaenoic acid—10.9 wt%. Despite these enhancements, the maximum relative lipid production was achieved at 21 degrees Celsius—19.1%.