Isolation and characterization of the PEP-phosphomutase and the phosphonopyruvate decarboxylase genes from the phosphinothricin tripeptide producer Streptomyces viridochromogenes Tü494.
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.
Eberhard-Karls-Universität Tübingen, Germany.
D
D
Schwartz
J
J
Recktenwald
S
S
Pelzer
W
W
Wohlleben
D000595:Amino Acid Sequence; D000613:Aminobutyrates / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D001426:Bacterial Proteins; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D015139:Blotting, Southern; D002262:Carboxy-Lyases / Q000235:genetics* / Q000378:metabolism; D015723:Gene Library; D008969:Molecular Sequence Data; D009154:Mutation; D010453:Peptide Synthases / Q000235:genetics / Q000378:metabolism; D010455:Peptides; D017875:Phosphotransferases (Phosphomutases) / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics*
D000613:Aminobutyrates; D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D010455:Peptides; C003121:phosphinothricin; D002262:Carboxy-Lyases; C113679:PPD protein, Streptomyces viridochromogenes; D017875:Phosphotransferases (Phosphomutases); C057167:phosphoenolpyruvate mutase; D010453:Peptide Synthases