Glucose-6-phosphate dehydrogenase partitioning in two-phase aqueous mixed (nonionic/cationic) micellar systems

Area of Science:

• Biochemistry
• Separation Science
• Surfactant Chemistry

Background:

• Glucose-6-phosphate dehydrogenase (G6PD) is crucial for NADPH production and nucleotide biosynthesis.
• G6PD is a valuable analytical reagent in various quantitative assays.
• Efficient purification strategies for G6PD are essential for its applications.

Purpose of the Study:

• To investigate the partitioning behavior of G6PD in two-phase aqueous mixed (nonionic/cationic) micellar systems.
• To explore new strategies for purifying G6PD using mixed micellar systems.
• To optimize the balance between enzyme stability and separation efficiency.

Main Methods:

• Experimental and theoretical investigation of G6PD partitioning in mixed micellar systems.
• Utilized two-phase aqueous systems composed of nonionic surfactant C(10)E(4) and cationic surfactants C(n)TAB (n=8, 10, 12).
• Assessed the effect of cationic surfactant tail length on G6PD denaturation and partitioning.

Main Results:

• Mixed micellar systems significantly improved G6PD partitioning compared to single-component systems.
• Electrostatic attractions between positively charged micelles and negatively charged G6PD drove preferential partitioning.
• The C(10)E(4)/C(10)TAB system yielded the highest partition coefficient (7.7) and G6PD yield (71%).
• C(8)TAB was least denaturing, but C(12)TAB provided stronger electrostatic attractions.

Conclusions:

• Two-phase aqueous mixed micellar systems offer an effective strategy for G6PD purification.
• The C(10)E(4)/C(10)TAB system provides an optimal balance for G6PD separation.
• Theoretical predictions of partition coefficients showed good agreement with experimental data.