Structural Identification of Autoinducer of Photobacterium Fischeri Luciferase

Biochemistry. 1981 Apr 28;20(9):2444-9. doi: 10.1021/bi00512a013.

Abstract

Synthesis of bacterial luciferase in some strains of luminous bacteria requires a threshold concentration of an autoinducer synthesized by the bacteria and excreted into the medium. Autoinducer excreted by Photobacterium fischeri strain MJ-1 was isolated from the cell-free medium by extraction with ethyl acetate, evaporation of solvent, workup with ethanol-water mixtures, and silica gel chromatography, followed by normal-phase and then reverse-phase high-performance liquid chromatography. The final product was greater than 99% pure. The structure of the autoinducer as determined by high-resolution 1H nuclear magnetic resonance spectroscopy, infrared spectroscopy, and high-resolution mass spectrometry was N-(3-oxohexanoly)-3-aminodihydro-2(3H)-furanone [or N-(beta-ketocaproyl)homoserine lactone]. The formation of homoserine by hydrolysis of the autoinducer was consistent with this structure. Synthetic autoinducer, obtained as a racemate, was prepared by coupling homoserine lactone to the ethylene glycol ketal of sodium 3-oxohexanoate, followed by mildly acidic removal of the protecting group; this synthetic material showed the appropriate biological activity.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 4-Butyrolactone / analogs & derivatives
  • 4-Butyrolactone / isolation & purification
  • 4-Butyrolactone / physiology*
  • Enzyme Induction
  • Furans / physiology*
  • Kinetics
  • Luciferases / biosynthesis*
  • Luminescent Measurements
  • Magnetic Resonance Spectroscopy
  • Mass Spectrometry
  • Photobacterium / enzymology*

Substances

  • Furans
  • N-(3-oxohexanoyl)-3-aminodihydro-2(3H)-furanone
  • Luciferases
  • 4-Butyrolactone