Chromochloris　zofingiensis,　capable　of　high-cell-density　heterotrophic　cultivation,　is　deemed　a　highly　promising　contender　for　producing　astaxanthin.　However,　the　relatively　low　astaxanthin　content　impedes　the　improvement　of　its　production.　Herein,　the　cultivation　conditions　for　heterotrophic　astaxanthin　production　were　optimized　in　this　study.　An　optimized　medium　was　proposed　with　a　culture　temperature　of　30　°C　and　an　initial　biomass　concentration　of　0.6　g/L.　Additionally,　a　two-stage　cultivation　strategy,　supplying　glucose　upon　carbon　depletion,　was　explored　to　enhance　both　biomass　concentration　and　astaxanthin　content.　Furthermore,　a　continuous　fed-batch　cultivation　strategy,　maintaining　the　glucose　concentration　at　around　5　g/L　under　nitrogen　deficiency,　was　established　in　a　5　L　fermenter.　This　approach　resulted　in　an　impressive　astaxanthin　content　of　1.92　mg/g　and　an　outstanding　astaxanthin　production　of　238.83　mg/L,　surpassing　most　reported　results　obtained　solely　through　fed-batch　operation　under　heterotrophic　cultivation.　The　findings　highlight　the　immense　potential　of　the　proposed　cultivation　strategy　for　commercial　astaxanthin　production　using　C.　zofingiensis.　©　2025　Elsevier　B.V.