Heterologous expression and localization of gentisate transporter Ncg12922 from Corynebacterium glutamicum ATCC 13032

Biochem Biophys Res Commun. 2006 Jul 28;346(2):555-61. doi: 10.1016/j.bbrc.2006.05.143. Epub 2006 Jun 2.


Ralstonia sp. strain U2 metabolizes naphthalene via gentisate (2,5-dihydroxybenzoate) to central metabolites, but it was found unable to utilize gentisate as growth substrate. A putative gentisate transporter encoded by ncg12922 from Corynebacterium glutamicum ATCC 13032 was functionally expressed in Ralstonia sp. strain U2, converting strain U2 to a gentisate utilizer. After ncg12922 was inserted into plasmid pGFPe with green fluorescence protein gene gfp, the expressed fusion protein Ncg12922-GFP could be visualized in the periphery of Escherichia coli cells under confocal microscope, consistent with a cytoplasmic membrane location. In contrast, GFP was ubiquitous in the cytoplasm of E. coli cells carrying pGFPe only. Gentisate 1,2-dioxygenase activity was present in the cell extract from strain U2 induced with gentisate but at a much lower level (one-fifth) than that obtained with salicylate. However, it exhibited a similar level in strain U2 containing Ncg12922 induced either by salicylate or gentisate.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Corynebacterium glutamicum / genetics
  • Corynebacterium glutamicum / metabolism*
  • Cytoplasm / metabolism
  • Dioxygenases / metabolism
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gentisates / metabolism*
  • Green Fluorescent Proteins / genetics
  • Membrane Transport Proteins / genetics
  • Membrane Transport Proteins / metabolism*
  • Ralstonia / genetics
  • Ralstonia / metabolism*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Salicylates / metabolism
  • Substrate Specificity


  • Bacterial Proteins
  • Gentisates
  • Membrane Transport Proteins
  • Recombinant Fusion Proteins
  • Salicylates
  • Green Fluorescent Proteins
  • Dioxygenases
  • gentisate 1,2-dioxygenase