In vitro Characterization of a Heterologously Expressed Non-Ribosomal Peptide Synthetase Involved in Phosphinothricin Tripepti

Synthetase Involved in Phosphinothricin Tripeptide Biosynthesis Jin-Hee Lee, Bradley S. Evans, Gongyong Li, Neil L. Kelleher, and Wilfred A. van der Donk SUPPORTING INFORMATION 1. Chemical Synthesis of Substrates Substrates. AcDMPT was synthesized using the radical addition of hypophosphorous acid to vinyl glycine as the key step as described previously (1) and similar to a procedure reported while this work was in progress (2). General All NMR spectra were recorded on Varian U500 spectrometers. H NMR spectra are referenced to TMS at 0 ppm or CHCl3 at 7.26 ppm, and C NMR spectra are referenced to CDCl3 at 77.7 ppm. Mass spectrometry (MS) experiments were carried out by the Mass Spectrometry Laboratory at the University of Illinois at Urbana-Champaign. Fractions collected during silica gel column chromatography were analyzed by TLC. Unless otherwise specified, all compounds and solvents were obtained from Fisher or Aldrich. THF was distilled from sodium/benzophenone, and CH2Cl2 was distilled form CaH2 prior to use. 1.1. Synthesis of AcDMPt.