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

Chem Commun (Camb). 2018 Oct 9;54(81):11467-11470. doi: 10.1039/c8cc06017g.

Abstract

Directed sortase A evolution yielded the variants R159G and D165Q/D186G/K196V with increased resistance (2.2-fold) and catalytic efficiency (6.3-fold) in 45% (v/v) dimethylsulfoxide. Interestingly, D165Q/D186G/K196V also showed an up to 4.7-fold increased activity for the conjugation of hydrophobic peptides/amines in co-solvents. MD simulations revealed that conformational mobilities are important for the gained resistance.

MeSH terms

  • Amino Acid Sequence
  • Aminoacyltransferases / chemistry*
  • Aminoacyltransferases / genetics
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Catalysis
  • Cysteine Endopeptidases / chemistry*
  • Cysteine Endopeptidases / genetics
  • Dimethyl Sulfoxide / chemistry
  • Dimethylformamide / chemistry
  • Directed Molecular Evolution
  • Ethanol / chemistry
  • Methanol / chemistry
  • Molecular Dynamics Simulation
  • Mutagenesis, Site-Directed
  • Mutation
  • Peptides / chemical synthesis
  • Peptides / chemistry*
  • Protein Engineering
  • Solvents / chemistry*
  • Staphylococcus aureus / enzymology
  • Water / chemistry

Substances

  • Bacterial Proteins
  • Peptides
  • Solvents
  • Water
  • Ethanol
  • Dimethylformamide
  • Aminoacyltransferases
  • sortase A
  • Cysteine Endopeptidases
  • Methanol
  • Dimethyl Sulfoxide