Enhanced macrocyclizing activity of the thioesterase from tyrocidine synthetase in presence of nonionic detergent. Academic Article uri icon

Overview

abstract

  • Macrocyclization carried out by thioesterase domains of multimodular nonribosomal peptide synthetases (NRPSs) is a key step in the biosynthesis of many biologically active peptides. The thioesterase excised from tyrocidine synthetase is a versatile macrocyclization catalyst and a useful tool for chemoenzymatic synthesis of diverse cyclic peptides. However, its utility is limited by its short lifetime of catalytic activity as well as significant flux of the acyl-enzyme intermediate to hydrolysis. The addition of Brij 58, a nonionic detergent, above the critical micelle concentration, has dramatic effects on enzyme activity: catalytic activity is extended to >60 min and the rate of cyclization (but not hydrolysis) increases 6-fold, resulting in a net 150- to 300-fold increase in cyclic product yields. This enhanced activity allowed enzymatic macrocyclization of a solid phase library of tyrocidine decapeptides to identify acceptable substitutions at the Orn9 position which had previously been inaccessible for diversification.

publication date

  • November 1, 2004

Research

keywords

  • Cetomacrogol
  • Detergents
  • Peptide Synthases
  • Thiolester Hydrolases

Identity

Scopus Document Identifier

  • 8844247217

Digital Object Identifier (DOI)

  • 10.1016/j.chembiol.2004.09.003

PubMed ID

  • 15556008

Additional Document Info

volume

  • 11

issue

  • 11