Designing biological fluid inspired molecularly crowded ionic liquid media as a sustainable packaging platform for cytochrome c.

Biological fluids are highly crowded due to the presence of various biomolecules. Therefore, it is essential to study the effects of molecular crowding agents to probe the behavior of a protein in a cell-like environment. Inspired by biofluids, herein, a molecularly crowded environment is created in the presence of an ionic liquid (IL), envisaging sustainable protein packaging. Interestingly, the molecularly crowded IL media enhanced the stability and catalytic activity (1.5-fold higher) of cytochrome c (Cyt c) as compared to the IL alone and the crowding agents without the IL. A similar trend was observed when the activity was recorded at 100 °C and when stored at room temperature for 30 days. Moreover, Cyt c dissolved in the molecularly crowded IL media was regenerated successfully without affecting the melting temperature of the protein, confirming the suitability of the molecularly crowded IL media as a potential and ecofriendly packaging system for Cyt c.