LAUSR

Abstract Ionic liquid-based aqueous biphasic system (ILABS) has emerged as an attractive green approach for the extraction and separation of various biomolecules. The growing market demands of cytochrome c (cyt c) due to its vast uses in medical applications have urged the search for cost-efficient approaches for the production and purification of cyt c. In this study, the feasibility of ILABS to recover cyt c from simulated Saccharomyces cerevisiae cell cultures was investigated by evaluating the effects of phase composition, pH, and additives concentration on the recovery efficiency of cyt c. The ILABS was developed using the hydrophilic ionic liquids, 1-hexyl-3-methylimidazolium bromide ((C6mim)Br) and potassium citrate. The optimal separation conditions for recovery of cyt c from simulated cells cultures were attained with ILABS of pH 9 comprising of 28% (w/w) of (C6mim)Br and 24% (w/w) of potassium citrate and the addition of 0.2% (w/w) NaCl at room temperature (25 °C). Cyt c was recovered in the IL-rich phase with partition coefficient (Kc) of 364.00 ± 1.09, recovery yield (Y) of 99.76% ± 0.03 and selectivity (S) of 223.31 ± 0.56. The results suggest that ILABS can efficiently recover cyt c from microbial fermentation broth with high recovery yields and separation efficiency.