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A split-gene transketolase essential for phosphonothrixin biosynthesis.

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This study identifies the phosphonothrixin (PTX) gene cluster and characterizes its biosynthetic enzymes. Researchers discovered a novel cooperative mechanism between PtxB5/6 and PtxB7, crucial for PTX formation.

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Area of Science:

  • Natural product biosynthesis
  • Enzymology
  • Actinobacteria metabolism

Background:

  • Phosphonothrixin (PTX) is a herbicidal compound produced by Saccharothrix sp. ST-888.
  • The biosynthesis of PTX involves a phosphonic acid moiety and an acetyl group.
  • A split-gene transketolase, PtxB5/6, was previously proposed to mediate acetylation.

Purpose of the Study:

  • To identify the PTX biosynthetic gene cluster in Saccharothrix sp. ST-888.
  • To characterize the enzymes involved in PTX biosynthesis.
  • To elucidate the reaction mechanism of PTX formation.

Main Methods:

  • Identification of the PTX biosynthetic gene cluster.
  • Cloning, expression, and purification of recombinant PtxB5/6.
  • In vitro enzymatic assays using purified proteins.

Main Results:

  • The PTX biosynthetic gene cluster was identified in Saccharothrix sp. ST-888.
  • Recombinant PtxB5/6 recognized hydroxyethyl-thiamine diphosphate as a substrate.
  • PtxB5/6 catalyzed the stereoselective acetylation of (3-hydroxy-2-oxopropyl)phosphonic acid, leading to PTX.

Conclusions:

  • A novel reaction mechanism for PTX biosynthesis was revealed.
  • The split-gene transketolase PtxB5/6 and acetohydroxyacid synthase homolog PtxB7 act cooperatively.
  • This cooperative action is essential for the formation of phosphonothrixin.