Enzyme encapsulation in permeabilized Saccharomyces cerevisiae cells

Area of Science:

• Biotechnology
• Cell Biology
• Biochemistry

Background:

• The Saccharomyces cerevisiae cell wall acts as a semipermeable barrier, controlling molecular passage.
• Enzymatic activity can be retained within cells after membrane extraction, with potential for reuse.

Purpose of the Study:

• To investigate the properties of permeabilized Saccharomyces cerevisiae cells as biocatalysts.
• To explore methods for modifying cell wall permeability and its impact on enzyme encapsulation and activity.

Main Methods:

• Detergent treatment to permeabilize Saccharomyces cerevisiae cells expressing E. coli lacZ.
• Modification of cell wall permeability through strain selection (Sigma1278b vs. BY4742/W303) and gene deletion (EXG1).
• Assessment of enzyme activity and reusability in permeabilized cells, including encapsulated enzyme systems for ATP regeneration.

Main Results:

• Permeabilized S. cerevisiae cells retained significant beta-galactosidase activity and could be reused over 15 times.
• Cell wall permeability varied between strains and could be reduced by deleting EXG1 or increased with Zymolyase treatment.
• Encapsulating multiple enzymes within permeabilized cells demonstrated kinetic advantages for ATP regeneration compared to cell extracts.

Conclusions:

• Permeabilized Saccharomyces cerevisiae cells serve as robust and reusable biocatalysts.
• Tailoring cell wall permeability through genetic modification or enzymatic treatment offers a strategy for optimizing biocatalytic systems.
• Enzyme encapsulation in permeabilized cells presents a promising approach for enhancing enzymatic reaction kinetics and stability.