Directed sortase A evolution for efficient site-specific bioconjugations in organic co-solvents

15/10/2018
 

Zhi Zou, Hoda Alibiglou, Diana M. Mate, Mehdi D. Davari, Felix Jakob, and Ulrich Schwaneberg, Chemical Communications, 2018, 54, 11467-11470

Evolved sortase A variants enable efficient site-specific conjugation of peptide/amine in organic co-solvents

 
  Photo of Zhi Zou Copyright: Bio VI

Evolved sortase A variants enable efficient site-specific conjugation of peptide/amine in organic co-solvents

Directed evolution of sortase A in 45% (v/v) DMSO, as co-solvent, yielded variants M1 and M3 with 2.2-fold increased resistance and 6.3-fold increased catalytic efficiency compared to sortase A wild-type, respectively. The generated variants were used to expand sortase-mediated ligation in organic co-solvent with hydrophobic substrates. Variant M3 showed up to 4.7-fold increased activity for peptide-peptide/peptide-amines in DMSO/DMF co-solvents compared to the wild-type. Structure-function relationship of sortase A in DMSO co-solvent were investigated by computational studies, which revealed that conformational mobility is important for the resistance gained by sortase A in organic co-solvent.

 
  Scheme Directed Evolution of Sortase A Copyright: Royal Society of Chemistry Schematic representation of sortase A -M3 mediated site-specific ligation in organic co-solvent

This work was financed by the Chinese Scholarship Council-CSC. Simulations were performed with computing resources granted by JARA-HPC from RWTH Aachen University under projects RWTH0116 and JARA0169.

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