Process‐Driven Acetate‐Based Lipid Production by the Oleaginous Yeast <em>Lipomyces starkeyi</em>

Oleaginous yeasts are promising microbial platforms for lipid production from non-conventional carbon sources; however, acetate utilization is frequently constrained by physiological limitations associated with culture pH. In this study, acetate utilization, biomass formation, and lipid production by Lipomyces starkeyi were investigated under flask and fed-batch cultivation to evaluate the influence of culture pH and pH control strategy. Statistically supported flask-scale experiments demonstrated that acetate concentration and cultivation time significantly affected acetate consumption, biomass formation, lipid yield, and culture pH, with excessive acetate loading resulting in culture alkalization, incomplete substrate utilization, and reduced process performance. Although lipid yield increased with increasing acetate concentration, lipid content and fatty acid composition remained unchanged, indicating that enhanced lipid production was primarily attributable to increased biomass formation rather than to changes in lipid biosynthesis. Fed-batch cultivation under different pH-control strategies provided qualitative insights into the relationships among pH regulation, acetate availability, and lipid accumulation under controlled fermentation conditions. While lipid accumulation was observed under both HCl-based and acetic acid–based pH control, differences in pH stability and cumulative acetate availability were associated with distinct patterns of lipid production. Collectively, these results identify culture pH as a critical physiological parameter influencing acetate utilization and lipid accumulation in L. starkeyi and suggest that coordinated pH control and carbon feeding strategies may improve the robustness of acetate-based lipid production processes. Further replicated fed-batch studies will be required to quantitatively validate these trends and support industrial applications.