Reverse micellar extraction of papain with cationic detergent based system: An optimization approach

Area of Science:

• Biochemistry
• Separation Science
• Chemical Engineering

Background:

• Reverse micellar extraction is a key technique for isolating biomolecules.
• Optimizing extraction efficiency (BEE) is crucial for cost-effective papain recovery.
• Cetyltrimethylammonium bromide (CTAB) is a common surfactant in reverse micellar systems.

Purpose of the Study:

• To optimize the forward and back extraction efficiency (BEE) of papain using a CTAB-based reverse micellar system.
• To determine the optimal conditions for papain extraction and purification.
• To improve the overall enzyme recovery and activity.

Main Methods:

• Reverse micellar extraction using cetyltrimethylammonium bromide (CTAB)/iso-octane/hexanol/butanol.
• Optimization of aqueous phase pH, CTAB concentration, and NaCl concentration for forward extraction.
• Application of Taguchi's orthogonal array to optimize stripping phase pH, isopropyl alcohol (IPA), and potassium chloride (KCl) concentrations.
• Further optimization using an artificial neural network-linked genetic algorithm.

Main Results:

• Maximum forward extraction efficiency achieved at pH 11.0, 150 mM CTAB, and 0.1 M NaCl.
• Optimal stripping conditions determined as pH 6, 20% IPA, and 0.8 M KCl, yielding 88% BEE.
• Artificial neural network-linked genetic algorithm further improved BEE to 90.52% under specific conditions.
• Enzyme activity was recovered with a 2.5-fold purification.

Conclusions:

• The study successfully optimized reverse micellar extraction for papain recovery.
• The developed method significantly enhances back extraction efficiency and enzyme purification.
• This approach offers a promising strategy for efficient and cost-effective papain isolation.