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Improving Sortase-A-Mediated Transpeptidation Reactions by Electrostatically Assisted Capture.

Chen Wang¹, Rémi Desmet², Benoît Snella²

¹Centrale Lille, F-59000 Lille, France.

Organic Letters

|October 9, 2025

Summary

This summary is machine-generated.

Researchers engineered Sortase A (SrtA) enzyme substrates by adding a polyarginine tag. This modification enhances protein modification yields by capturing released peptides, improving SrtA efficiency.

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Area of Science:

Biochemistry
Protein Engineering
Enzymology

Background:

Sortase A (SrtA) is a transpeptidase crucial for protein modification.
The reversible nature of SrtA reactions limits product yields.
Developing strategies to enhance SrtA efficiency is important for protein engineering.

Purpose of the Study:

To engineer Sortase A substrates to improve reaction yields.
To develop a method for sequence-specific capture of released peptides.
To enhance the efficiency of SrtA-catalyzed transpeptidation.

Main Methods:

Engineered a novel substrate by extending the LPxTG recognition motif with a polyarginine peptide module.
Utilized electrostatically assisted aminolysis for sequence-specific peptide capture.
Employed a negatively charged thioester as the nucleophile.

Main Results:

The engineered substrate enabled sequence-specific capture of the released peptide.
The strategy effectively shifted the reaction equilibrium toward product formation.
Achieved improved SrtA efficiency under mild conditions with moderate nucleophile excess.

Conclusions:

Substrate engineering with a polyarginine tag is a viable strategy to enhance SrtA activity.
Electrostatically assisted aminolysis provides a traceless and selective method for product isolation.
This approach offers a significant improvement for SrtA-mediated protein modification.