Abstract Objectives: Natural antioxidants particularly carotenoids have been associated with a lowered risk for stroke and cerebrovascular diseases. The present study proposes a new microbial source of carotenoid-rich bio-pigment and investigates its potency in mitigating ischemic stroke in an animal model. Methods:__ampersandsignnbsp; A yeast isolate rich in carotenoid pigment was defined morphologically and physiologically, recognized by 18S rDNA as Rhodotorula mucilaginosa G20with a similarity of 100%, then submitted to Gen Bank (accession number KY271337.1). The extracted pigment was analyzed using high-performance liquid chromatography invitro.__ampersandsignnbsp; Furthermore, an in-vivo rat model was assessed to investigate the effect of the extracted pigment against Endothelin-1 (ET-1) induced focal cerebral ischemia. Rats were assigned into four groups: Group (1) normal control rats received saline. Group (2): sham-operated rats received saline, Group (3): rats received ET-1.Group (4): rats received extracted pigment and ET.__ampersandsignnbsp; Results: HPLC results revealed that the extracted yeast pigment consists mainly of neoxanthin and __ampersandsignbeta;-carotene. Invivo, pretreatment with extracted pigment significantly enhanced the grip strength, restored impaired vertical forelimb use induced by ET-1__ampersandsignnbsp; in rats, halted oxidants biomarkers, and triggered antioxidant mediators. Moreover, it suppressed inflammatory and apoptotic markers. The histological and structural deterioration in the ventral subiculum was deterred. Conclusions: Based on these encouraging results, The extracted microbial carotenoid pigment production can be considered efficient and economical and proves to be a promising approach for ameliorating cerebral ischemic injuries.