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
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.
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.
