Sequence analysis and functional characterization of the violacein biosynthetic pathway from Chromobacterium violaceum.
Violacein is a purple-colored, broad-spectrum antibacterial pigment that has a dimeric structure composed of 5-hydroxyindole, oxindole and 2-pyyrolidone subunits formed by the condensation of two modified tryptophan molecules. The violacein biosynthetic gene cluster from Chromobacterium violaceum was characterized by DNA sequencing, transposon mutagenesis, and chemical analysis of the pathway intermediates produced heterologously in Escherichia. coli. The violacein biosynthetic gene cluster spans eight kilobases and is comprised of the four genes, vioABCD, that are necessary for violacein production. Sequence analysis suggests that the products of vioA, vioC and vioD are nucleotide-dependent monooxygenases. Disruption of vioA or vioB completely abrogates the biosynthesis of violacein intermediates, while disruption of the vioC or vioD genes results in the production of violacein precursors.
August
P R
PR
Ariad Pharmaceuticals, Inc, Cambridge, Massachusetts 02139, USA. Paul.august@aventis.com
Grossman
T H
TH
Minor
C
C
Draper
M P
MP
MacNeil
I A
IA
Pemberton
J M
JM
Call
K M
KM
Holt
D
D
Osburne
M S
MS
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents; D001483:Base Sequence; D002861:Chromobacterium / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli; D007211:Indoles / Q000378:metabolism*; D006899:Mixed Function Oxygenases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D005810:Multigene Family; D016254:Mutagenesis, Insertional; D009876:Operon; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014364:Tryptophan / Q000378:metabolism
D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D007211:Indoles; D014364:Tryptophan; D006899:Mixed Function Oxygenases; C063155:violacein