Isolation and characterization of the PEP-phosphomutase and the phosphonopyruvate decarboxylase genes from the phosphinothricin tripeptide producer Streptomyces viridochromogenes Tü494

FEMS Microbiol Lett. 1998 Jun 15;163(2):149-57. doi: 10.1111/j.1574-6968.1998.tb13039.x.

Abstract

The previously isolated non-phosphinothricin tripeptide producing Streptomyces viridochromogenes gene disruption mutant SP62/2 was used to identify and analyze genes encoding early steps of the phosphinothricin tripeptide biosynthesis. Cross-feeding and bioconversion experiments between SP62/2 and known non-phosphinothricin tripeptide producing mutants or presumptive phosphinothricin tripeptide precursors revealed that SP62/2 was blocked in step one or two of the phosphinothricin tripeptide biosynthesis. It was shown that the block in the biosynthesis is due to the integration of a temperature-sensitive plasmid by illegitimate recombination into the phosphinothricin tripeptide biosynthetic gene cluster. The corresponding region was isolated from the wild-type. A 2.7-kb DNA fragment was analyzed comprising three ORFs (ppm, ppd, orfX) which are probably translationally coupled. The ppm gene encodes a protein which is similar to PEP-phosphomutases and the deduced Ppd product shows similarity to the phosphonopyruvate decarboxylase from Streptomyces wedmorensis.

MeSH terms

  • Amino Acid Sequence
  • Aminobutyrates / metabolism*
  • Anti-Bacterial Agents / biosynthesis*
  • Bacterial Proteins*
  • Bacteriophage lambda / genetics
  • Base Sequence
  • Blotting, Southern
  • Carboxy-Lyases / genetics*
  • Carboxy-Lyases / metabolism
  • Gene Library
  • Molecular Sequence Data
  • Mutation
  • Peptide Synthases / genetics
  • Peptide Synthases / metabolism
  • Peptides*
  • Phosphotransferases (Phosphomutases) / genetics*
  • Phosphotransferases (Phosphomutases) / metabolism
  • Plasmids / genetics
  • Sequence Analysis, DNA
  • Streptomyces / enzymology
  • Streptomyces / genetics*

Substances

  • Aminobutyrates
  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Peptides
  • phosphinothricin
  • Carboxy-Lyases
  • PPD protein, Streptomyces viridochromogenes
  • Phosphotransferases (Phosphomutases)
  • phosphoenolpyruvate mutase
  • Peptide Synthases

Associated data

  • GENBANK/Y14337