In vitro characterization of a heterologously expressed nonribosomal peptide synthetase involved in phosphinothricin tripeptide biosynthesis

Jin Hee Lee, Bradley S. Evans, Gongyong Li, Neil L. Kelleher, Wilfred A. Van Der Donk

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

The late stages of biosynthesis of phosphinothricin tripeptide (PTT) involve peptide formation and methylation on phosphorus. The exact timing of these transformations is not known. To provide insight into this question, we developed a heterologous expression system for PhsA, one of three NRPS proteins in PTT biosynthesis. The apparent kcat/Km value for ATP-pyrophosphate exchange activity for D,L-N-acetylphosphinothricin was 3.5 μM-1 min-1, whereas the kcat/K m,app for L-N-acetyldemethylphosphinothricin was 0.5 μM -1 min-1, suggesting the former might be the physiological substrate. Each substrate could be loaded onto the phosphopantetheine arm of the thiolation domain as observed by Fourier transform mass spectrometry (FTMS).

Original languageEnglish (US)
Pages (from-to)5054-5056
Number of pages3
JournalBiochemistry
Volume48
Issue number23
DOIs
StatePublished - Jun 16 2009

ASJC Scopus subject areas

  • Biochemistry

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