Mapping of endoglucanases displayed on yeast cell surface using atomic force microscopy

Area of Science:

• Biotechnology
• Biophysics
• Surface Science

Background:

• Yeast cell surfaces offer potential for cellulose utilization.
• Visualizing and evaluating surface-displayed enzymes like cellulase remains challenging.

Purpose of the Study:

• To develop and demonstrate a method for visualizing endoglucanase (EG) on yeast cell surfaces.
• To investigate the impact of different anchoring regions on EG display and localization.
• To measure the interactive forces between cellulose and surface-displayed EG.

Main Methods:

• Utilized two unique anchoring regions for displaying endoglucanase (EG) on yeast cell surfaces.
• Employed atomic force microscopy (AFM) to observe EG display levels and localization.
• Used chemically modified AFM cantilevers to measure the interactive force between cellulose and EG.
• Performed force curve mapping to analyze EG distribution and interaction.

Main Results:

• Observed distinct differences in EG display levels and localization based on the anchoring regions used.
• Quantified the interactive forces between cellulose and surface-displayed EG.
• Force curve mapping confirmed variations in EG display and localization.

Conclusions:

• The proposed methodology effectively visualizes displayed enzymes, such as endoglucanase, on yeast cell surfaces.
• Anchoring regions significantly influence the display and localization of EG.
• AFM provides a powerful tool for evaluating surface-displayed enzyme interactions.