Integrated Approach to Extract and Purify Proteins from Honey by Ionic Liquid-Based Three-Phase Partitioning

Area of Science:

• Biotechnology
• Green Chemistry
• Biochemistry

Background:

• Three-phase partitioning (TPP) offers efficient compound purification from liquid-liquid interphases.
• Conventional TPP methods often require high salt and solvent concentrations, impacting sustainability.
• Developing greener TPP systems is crucial for environmentally friendly bioprocessing.

Purpose of the Study:

• To investigate the use of biobased ionic liquids (ILs) and honey in TPP systems for sustainable protein purification.
• To apply these novel IL-TPP systems for the purification of major royal jelly proteins (MRJPs) from honey.
• To assess the efficiency and sustainability of the proposed IL-TPP approach.

Main Methods:

• Development of TPP systems using biobased glycine-betaine ionic liquids and honey as phase-forming components.
• Application of the IL-TPP systems to the purification of MRJPs from honey.
• Quantification of recovery yields and purity levels of target proteins and other compounds.

Main Results:

• MRJPs predominantly precipitated in the IL-TPP interphase, achieving recovery yields from 82.8% to 97.3%.
• High purity levels, up to 90.1%, were obtained for MRJP1.
• Simultaneous separation of antioxidants and carbohydrates into different liquid phases was achieved.

Conclusions:

• The developed IL-TPP systems provide a sustainable and efficient method for purifying MRJPs from honey.
• This one-step process effectively separates proteins, antioxidants, and carbohydrates using only ILs and a natural matrix.
• The approach significantly improves the sustainability of TPP processes by minimizing salt and solvent usage.