Development of deep eutectic solvent-based aqueous biphasic system for the extraction of lysozyme

Area of Science:

• Biochemistry
• Separation Science
• Materials Science

Background:

• Deep eutectic solvents (DESs) offer tunable properties for separation processes.
• Aqueous biphasic systems (ABSs) are effective for biomolecule extraction.
• Lysozyme extraction from natural sources requires efficient and biocompatible methods.

Purpose of the Study:

• To develop and optimize DES-based ABS for lysozyme extraction from chicken egg white.
• To investigate the influence of DES structure and operational parameters on extraction efficiency.
• To evaluate the biocompatibility and mechanism of the lysozyme extraction process.

Main Methods:

• Synthesis and characterization of six novel DESs using quaternary ammonium salts and carboxylic acids.
• Evaluation of phase-formation ability of DESs with various salts (e.g., Na2SO4).
• Optimization of extraction parameters including DES/salt concentration, temperature, pH, and ionic strength.
• Analysis of lysozyme activity and structural integrity using spectroscopic and microscopic techniques.

Main Results:

• The [TBAB][Gly] and Na2SO4 system demonstrated efficient phase separation and lysozyme extraction.
• Over 98% lysozyme transfer into the DES-rich phase was achieved under optimal conditions.
• Lysozyme retained 91.73% of its initial activity, indicating high biocompatibility.
• The method was successfully applied to real chicken egg white samples.

Conclusions:

• DES-based ABS provide a highly efficient and biocompatible platform for lysozyme extraction.
• The study elucidates the key factors influencing phase separation and extraction efficiency.
• The developed method holds significant promise for the analysis and purification of biological samples.