Work

In Vitro Characterization of a Heterologously Expressed Nonribosomal Peptide Synthetase Involved in Phosphinothricin Tripeptide Biosynthesis

Year: 2009

Type: article

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 k(cat)/K(m) value for ATP-pyrophosphate exchange activity for d,l-N-acetylphosphinothricin was 3.5 muM(-1) min(-1), whereas the k(cat)/K(m,app) for l-N-acetyldemethylphosphinothricin was 0.5 microM(-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).

Source: Biochemistry

DOI: 10.1021/bi900164d

Authors: Jin-Hee Lee, Bradley S. Evans, Gongyong Li, Neil L. Kelleher, Wilfred A. van der Donk

Institutions: Carl R. Woese Institute for Genomic Biology, University of Illinois Urbana-Champaign, Howard Hughes Medical Institute

Language: English

Topic: Microbial Natural Products and Biosynthesis

Subfield: Pharmacology (medical)

Field: Medicine

Domain: Health Sciences

Open Access status: hybrid

Estimated APC paid: $4,500

Funders: Howard Hughes Medical Institute, National Institutes of Health, National Institute of General Medical Sciences