Chitosan microspheres as immobilized dye affinity support for catalase adsorption

Area of Science:

• Biomaterials Science
• Biochemistry
• Separation Science

Background:

• Chitosan microspheres (CS) are versatile biomaterials.
• Dye-ligand chromatography is a common protein purification technique.
• Catalase (CAT) is an important enzyme with various applications.

Purpose of the Study:

• To develop a novel dye-affinity adsorbent for catalase (CAT) adsorption.
• To investigate the adsorption capacity and conditions for CAT using the developed adsorbent.
• To evaluate the efficiency and non-denaturing properties of the adsorbent for CAT elution.

Main Methods:

• Preparation of chitosan microspheres (CS) via phase-inversion.
• Covalent attachment of Cibacron Blue F3GA (CB) to CS to form CB-CS adsorbent.
• Adsorption studies of CAT onto CB-CS, including capacity determination at varying pH.
• Application of Langmuir adsorption model.
• Elution studies using different conditions to assess recovery and denaturation.

Main Results:

• The maximum CAT adsorption capacity of CB-CS was 28.4 mg/g at pH 7.0.
• The Langmuir adsorption model accurately described the CAT adsorption process.
• Up to 90.6% of adsorbed CAT was successfully eluted using 0.5 M NaSCN at pH 8.0.
• The elution process did not cause significant denaturation of the adsorbed CAT.

Conclusions:

• CB-CS is a highly effective adsorbent for CAT.
• The developed adsorbent offers a high adsorption capacity and efficient elution.
• CB-CS shows potential for the purification of CAT without denaturation, making it suitable for biotechnological applications.