Highly efficient method towards in situ immobilization of invertase using cryogelation

Area of Science:

• Biotechnology
• Enzyme immobilization
• Biocatalysis

Background:

• Enzyme immobilization is crucial for industrial biocatalysis, improving enzyme stability and reusability.
• Developing efficient immobilization techniques for enzymes like Saccharomyces cerevisiae invertase is essential for applications such as invert sugar production.

Purpose of the Study:

• To develop a novel method for immobilizing Saccharomyces cerevisiae invertase within a supermacroporous polyacrylamide cryogel.
• To evaluate the efficiency, stability, and kinetic properties of the immobilized invertase for invert sugar production.

Main Methods:

• Enzyme cross-linking with polyglutaraldehyde (PGA) prior to immobilization to prevent leakage.
• In situ immobilization via free radical polymerization in a semi-frozen state within the cryogel matrix.
• Characterization of immobilized enzyme activity, stability, optimum conditions, and kinetic parameters.

Main Results:

• Achieved 100% immobilization and 74% activity yield.
• Immobilized invertase retained full activity for 30 days and 30 batch reactions.
• Immobilized enzyme showed enhanced stability at high pH and temperatures, with a shift in optimum pH from 4.5 to 5.0.

Conclusions:

• The developed cryogel immobilization method is simple, effective, and enhances enzyme stability.
• This technique shows potential for industrial applications in invert sugar production and immobilization of other enzymes/microorganisms.