Extraction of ascorbate oxidase from Cucurbita maxima by continuous process in perforated rotating disc contactor...

Area of Science:

• Biochemistry
• Biotechnology
• Separation Science

Background:

• Ascorbate oxidase is crucial for ascorbic acid analysis in pharmaceuticals, food, and clinics.
• Current purification methods for ascorbate oxidase are costly.
• Developing cost-effective purification techniques is essential.

Purpose of the Study:

• To investigate the continuous pre-purification of ascorbate oxidase using an aqueous two-phase system (PEG/citrate).
• To optimize conditions for efficient enzyme separation in a rotating column system.

Main Methods:

• Utilized an aqueous two-phase system composed of polyethylene glycol (PEG) and citrate.
• Employed a rotating column with perforated discs for continuous separation.
• Evaluated PEG molar mass, PEG concentration, and citrate concentration as key parameters.

Main Results:

• Optimal conditions identified: 20,000 g/mol PEG molar mass, 10% PEG concentration, and 25% citrate concentration.
• Achieved a partition coefficient of 3.35.
• Demonstrated a separation efficiency of 54.98% and a purification factor of 1.46.

Conclusions:

• The PEG/citrate aqueous two-phase system is suitable for the continuous pre-purification of ascorbate oxidase.
• This method offers a potentially cost-effective alternative to traditional enzyme purification techniques.
• The established conditions provide a foundation for scaling up the process for industrial applications.