Invertase immobilization via its carbohydrate moiety

Area of Science:

• Biochemistry
• Biotechnology
• Enzyme Engineering

Background:

• Enzyme immobilization is crucial for enzyme reusability and stability in industrial processes.
• Invertase, a key enzyme in sucrose hydrolysis, requires efficient immobilization strategies for practical applications.

Purpose of the Study:

• To investigate covalent immobilization of invertase onto various solid supports.
• To evaluate different immobilization methods, including direct coupling and modified Ugi reaction.
• To determine the impact of immobilization parameters on the activity of the resulting enzyme conjugate.

Main Methods:

• Periodate oxidation of invertase's glycosidic component to generate aldehyde groups.
• Covalent immobilization onto glycidyl methacrylate, styrene-divinylbenzene copolymers, and bead cellulose supports.
• Utilized direct reaction between enzyme aldehyde groups and support amino groups, and the modified Ugi reaction.

Main Results:

• Investigated effects of buffer, support type, oxidation conditions, and spacer length on enzyme activity.
• The modified Ugi reaction proved more effective for immobilization compared to direct coupling.
• Superior enzyme conjugate activity was achieved by immobilizing oxidized invertase onto styrene-divinylbenzene copolymers with amino groups.

Conclusions:

• The modified Ugi reaction offers an efficient method for invertase immobilization.
• Styrene-divinylbenzene copolymers with amino groups are optimal supports for enhanced invertase activity.
• Optimized immobilization conditions are critical for maximizing enzyme conjugate performance.