Integrating C-Phycocyanin, and Polyhydroxybutyrate Recovery Using a Triphasic System: Experimental Design and Optimization in Thermotolerant Potamosiphon sp.

This research assesses a triphasic extraction technique for the sequential retrieval of C-phycocyanin (C-PC) and polyhydroxybutyrate (PHB) from a thermotolerant Potamosiphon sp. strain. A two-stage design-of-experiments methodology was employed (Minimum Run Resolution V factorial design involving six variables, followed by a central composite design (CCD)) to optimize the chosen region. In the factorial stage, PHB ranged from 109.396 to 168.995 mg/g, and the model was significant (F = 22.63, p < 0.0001). Freeze-milling and vortexing were identified as critical elements, underscoring the importance of the t-butanol × (NH4)2SO4 interaction for phase selectivity. The CCD concentrating on freeze-milling and vortex cycles yielded a robust quadratic model (F = 78.18, p < 0.0001), forecasting a peak PHB yield of 191.82 mg/g at six freeze-milling cycles and three vortex cycles (desirability 0.921), while maintaining t-butanol at 19.9 mL, t-butanol concentration at 94.7% (v/v), (NH4)2SO4 at 49.9% (w/v), and vortex duration at 1.2 min. Ten separate trials validated the model’s accuracy, yielding an observed PHB of 191.5 mg/g, which closely matched the model’s prediction. The platform facilitates an integrated downstream process in which C-PC is recovered under moderate conditions before triphasic partitioning. This enables the simultaneous valorization of pigment, lipophilic fraction, and biopolymer inside a unified cyanobacterial biorefinery process.