A newly isolated red-pigmented yeast, Sporobolomyces reniformis EMCC1691, was evaluated for its biotechnological potential in an integrated case study aimed at developing an efficient microbial cell factory for the valorization of delactosed whey. Fermentation trials in 5 L bioreactors demonstrated robust yeast growth on this dairy by-product, with complete consumption of glucose (21.86 g/L) and galactose (20.36 g/L), leading to the accumulation of approximately 6172 mg/L of lipids and 5634 µg/L of total carotenoids. Fatty acid analysis revealed a final concentration of 3924 mg/L, mainly represented by oleic (2037 mg/L), palmitic (779 mg/L), stearic (403 mg/L), and linoleic (362 mg/L) acids. HPLC analysis showed a pigment profile dominated by torularhodin, torulene, γ-carotene, and β-carotene. To complement downstream processing, the fermented culture was spray-dried into a stable powder and subsequently subjected to a simple, cost-effective, and unconventional mechanical pretreatment using a hydraulic press. This post-drying operation ensured extensive cell-wall disruption without the use of chemical agents or specialized equipment, thereby significantly enhancing the recoverability of intracellular lipids and carotenoids through supercritical CO₂ extraction. Under optimized conditions, SFE-CO₂ with ethanol recovered 92.18 ± 1.61 µg/g of total carotenoids, achieving an extraction efficiency of 84% relative to organic solvent extraction (109.17 ± 2.10 µg/g). Importantly, fermentation also reshaped the fatty acid composition of delactosed whey, shifting it toward a profile enriched in monounsaturated and polyunsaturated fatty acids, thereby further highlighting the metabolic impact and bioconversion potential of S. reniformis EMCC1691. Overall, this work highlights the technological relevance of a recently characterized yeast species and its potential to convert dairy by-products into high-value compounds within a proof-of-concept microbial cell factory framework, paving the way for future scale-up investigations. © 2026 by the authors.

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