Formation of indigo and related compounds from indolecarboxylic acids by aromatic acid-degrading bacteria: chromogenic reactions for cloning genes encoding dioxygenases that act on aromatic acids.
The p-cumate-degrading strain Pseudomonas putida F1 and the m- and p-toluate-degrading strain P. putida mt-2 transform indole-2-carboxylate and indole-3-carboxylate to colored products identified here as indigo, indirubin, and isatin. A mechanism by which these products could be formed spontaneously following dioxygenase-catalyzed dihydroxylation of the indolecarboxylates is proposed. Indolecarboxylates were employed as chromogenic substrates for identifying recombinant bacteria carrying genes encoding p-cumate dioxygenase and toluate dioxygenase. Dioxygenase gene-carrying bacteria could be readily distinguished as dark green-blue colonies among other colorless recombinant Escherichia coli colonies on selective agar plates containing either indole-2-carboxylate or indole-3-carboxylate.
Eaton
R W
RW
Gulf Ecology Division, U.S. Environmental Protection Agency, Gulf Breeze, Florida 32561, USA.
Chapman
P J
PJ
D000596:Amino Acids / Q000378:metabolism*; D002264:Carboxylic Acids; D002863:Chromogenic Compounds / Q000378:metabolism*; D003001:Cloning, Molecular / Q000379:methods*; D004396:Coloring Agents / Q000378:metabolism; D007203:Indigo Carmine; D007211:Indoles / Q000378:metabolism; D007510:Isatin / Q000378:metabolism; D010105:Oxygenases / Q000235:genetics*; D016958:Pseudomonas putida / Q000235:genetics* / Q000378:metabolism
D000596:Amino Acids; D002264:Carboxylic Acids; D002863:Chromogenic Compounds; D004396:Coloring Agents; D007211:Indoles; C058980:indole-2-carboxylic acid; C012382:indole-3-carboxylic acid; D007510:Isatin; D007203:Indigo Carmine; D010105:Oxygenases; C027185:indirubin