Biosynthesis of the 3,4-dihydroxybenzoate moieties of petrobactin by Bacillus anthracis.
The biosynthesis of the 3,4-dihydroxybenzoate moieties of the siderophore petrobactin, produced by B. anthracis str. Sterne, was probed by isotopic feeding experiments in iron-deficient media with a mixture of unlabeled and D-[(13)C6]glucose at a ratio of 5:1 (w/w). After isolation of the labeled siderophore, analysis of the isotopomers was conducted via one-dimensional (1)H and (13)C NMR spectroscopy, as well as (13)C-(13)C DQFCOSY spectroscopy. Isotopic enrichment and (13)C-(13)C coupling constants in the aromatic ring of the isolated siderophore suggested the predominant route for the construction of the carbon backbone of 3,4-DHB (1) involved phosphoenol pyruvate and erythrose-4-phosphate as ultimate precursors. This observation is consistent with that expected if the shikimate pathway is involved in the biosynthesis of these moieties. Enrichment attributable to phosphoenol pyruvate precursors was observed at C1 and C6 of the aromatic ring, as well as into the carboxylate group, while scrambling of the label into C2 was not. This pattern suggests 1 was biosynthesized from early intermediates of the shikimate pathway and not through later shikimate intermediates or aromatic amino acid precursors.
Koppisch
Andrew T
AT
Bioscience Division, Los Alamos National Laboratory, Los Alamos, New Mexico 87545, USA. koppisch@lanl.goV
Hotta
Kinya
K
Fox
David T
DT
Ruggiero
Christy E
CE
Kim
Chu-Young
CY
Sanchez
Timothy
T
Iyer
Srinivas
S
Browder
Cindy C
CC
Unkefer
Pat J
PJ
Unkefer
Clifford J
CJ
D001408:Bacillus anthracis / Q000737:chemistry* / Q000378:metabolism*; D001549:Benzamides / Q000737:chemistry* / Q000378:metabolism*; D062385:Hydroxybenzoates / Q000737:chemistry* / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D012765:Shikimic Acid / Q000737:chemistry
C520290:3,4-dihydroxybenzoate; D001549:Benzamides; D062385:Hydroxybenzoates; C450699:petrobactin; D012765:Shikimic Acid