Characterization and analysis of early enzymes for petrobactin biosynthesis in Bacillus anthracis.

Abstract

Recently, iron acquisition and, more specifically, enzymes involved in siderophore biosynthesis have become attractive targets for discovery of new antibiotics. Accordingly, targeted inhibition of the biosynthesis of petrobactin, a virulence-associated siderophore encoded by the asb locus in Bacillus anthracis, may hold promise as a potential therapy against anthrax. This study describes the biochemical characterization of AsbC, the first reported 3,4-dihydroxybenzoic acid-AMP ligase, and a key component in the biosynthesis of DHB-spermidine (DHB-SP), the first isolable intermediate in petrobactin biosynthesis. AsbC catalyzes adenylation to the corresponding AMP ester of the unusual precursor 3,4-dihydroxybenzoate, in addition to benzoate substrates bearing hydrogen bond-donating substituents at the para and meta positions on the phenyl ring. In a second reaction, AsbC catalyzes transfer of the activated starter unit to AsbD, an aryl carrier protein similar to acyl and peptidyl carrier proteins that function in fatty acid, polyketide, and nonribosomal peptide biosynthesis. A third protein, AsbE, is shown to be responsible for condensation of 3,4-dihydroxybenzoyl-AsbD with spermidine, providing the DHB-spermidine arms that are linked to citrate for assembly of petrobactin. On the basis of the selective substrate profile of AsbC, a nonhydrolyzable analogue of 3,4-DHB-AMP was synthesized and shown to effectively inhibit AsbC function in vitro.

• Authors: Brian F Pfleger, Jung Yeop Lee, Ravindranadh V Somu, Courtney C Aldrich, Philip C Hanna, David H Sherman
• Journal: Biochemistry (Biochemistry) Vol. 46 Issue 13 Pg. 4147-57 (Apr 03 2007) ISSN: 0006-2960 [Print] United States
• PMID: 17346033 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• Benzamides
• Carrier Proteins
• petrobactin
• 3,4-dihydroxybenzoic acid-AMP ligase, Bacillus anthracis
• Nucleotidyltransferases

Topics

• Bacillus anthracis (enzymology, metabolism)
• Benzamides (metabolism)
• Carrier Proteins (isolation & purification, metabolism)
• Cloning, Molecular
• Nucleotidyltransferases (antagonists & inhibitors, isolation & purification, metabolism)
• Substrate Specificity