A growth selection system for sucrose synthases (SuSy): design and test

Area of Science:

• Biocatalysis
• Enzyme Engineering
• Synthetic Biology

Background:

• High throughput screening (HTS) is crucial for developing industrial biocatalysts.
• Current methods for many enzymes, including sucrose synthase, are limited by throughput or cost.
• Robust biocatalysts are needed for sustainable, biobased chemical synthesis.

Purpose of the Study:

• To present a novel, high throughput selection system for sucrose synthase variants.
• To couple sucrose synthase activity directly to cellular growth for simplified screening.
• To facilitate the development of improved enzymes for industrial applications.

Main Methods:

• Developed a selection system linking sucrose synthase activity to host cell growth.
• Utilized a straightforward assay amenable to high throughput screening.
• Focused on sucrose synthase (SS) as a model enzyme class.

Main Results:

• Demonstrated a functional coupling between sucrose synthase activity and cell viability.
• Established a system enabling rapid screening of a large number of enzyme variants.
• Showcased the potential for identifying robust sucrose synthase variants.

Conclusions:

• The presented system offers a straightforward and high throughput approach for screening sucrose synthase.
• This method can accelerate the discovery of improved biocatalysts for industrial glycosylation.
• The approach has the potential to reduce costs and improve efficiency in biobased synthesis.