Antibiotic Action of
            <i>N</i>
            -Methylthioformohydroxamate Metal Complexes

Bell, Stuart; Friedman, Stanley; Leong, John M.

doi:10.1128/aac.15.3.384

Cited by 18 publications

(8 citation statements)

References 19 publications

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“…Where specified, M9 and TMS were supplemented with casamino acids (0.3%), tryptophan (0.003%), thiamine (0.002%), and glucose (0.2%) (CM9 and CTMS). Stock solutions of casamino acids were extracted with 8-hydroxyquinoline and chloroform (4) to remove contaminating iron. Crystal violet-pectate (CVP) agar (16) and pectate agar (5) were the selective media for culture of E. carotovora.…”

Section: Methodsmentioning

confidence: 99%

High-affinity iron uptake systems present in Erwinia carotovora subsp. carotovora include the hydroxamate siderophore aerobactin

Ishimaru

Loper

1992

J Bacteriol

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The phytopathogenic bacterium Erwinia carotovora subsp. carotovora W3C105 produced the hydroxamate siderophore aerobactin under iron-limiting conditions. A survey of 22 diverse strains of E. carotovora revealed that strain W3C105 alone produced aerobactin. The ferric-aerobactin receptor of strain W3C105 was an 80-kDa protein, identified by immunoblots of Sarkosyl-soluble proteins obtained from E. carotovora cells grown in iron-depleted medium and probed with antiserum raised against the 74-kDa ferric-aerobactin receptor encoded by the pColV-K30 plasmid of Escherichia coli. Genes determining aerobactin biosynthesis and uptake were localized to an 11.3-kb EcoRI-HindIII chromosomal fragment of strain W3C105. A 10-kb subclone of the fragment conferred on E. coli DH5 alpha both aerobactin biosynthesis and uptake, determined by cloacin DF13 sensitivity, the presence of the 80-kDa receptor protein, and iron-independent growth of E. coli clones. The aerobactin biosynthesis genes of E. carotovora W3C105 hybridized to those of the pColV-K30 plasmid of E. coli, but the restriction patterns of the aerobactin regions of E. coli and E. carotovora differed. Although the aerobactin region of enteric bacteria is commonly flanked by IS1-like sequences, IS1 sequences were not detected in the genomic DNA or the cloned aerobactin region of E. carotovora. E. coli DH5 alpha cells harboring cloned aerobactin biosynthesis genes from E. carotovora W3C105 produced greater quantities of aerobactin and the 80-kDa ferric-aerobactin receptor when grown in iron-limited than in iron-replete medium. Strain W3C105 grew on an iron-limited medium, whereas derivatives that lacked a functional aerobactin iron acquisition system did not grow on the medium. These results provide evidence for the occurrence and heterogeneity of aerobactin as a high-affinity iron uptake system of both clinical and phytopathogenic species of the Enterobacteriaceae. Although future studies may reveal a role for aerobactin in the virulence or ecology of strain W3C105, a functional aerobactin iron acquisition system is not necessary for the pathogenicity of E. carotovora.

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Section: Methodsmentioning

confidence: 99%

High-affinity iron uptake systems present in Erwinia carotovora subsp. carotovora include the hydroxamate siderophore aerobactin

Ishimaru

Loper

1992

J Bacteriol

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“…To avoid contamination with iron, glassware was cleaned with 6 M HCl (32). Casamino Acids were deferrated by extracting stock solutions with 3% (wt/wt) 8-hydroxyquinoline in chloroform (8). Water purified by reverse osmosis and by Milli-Q treatment (Millipore Corp., Bedford, Mass.)…”

Section: Methodsmentioning

confidence: 99%

Iron-Binding Compounds from Agrobacterium spp.: Biological Control Strain Agrobacterium rhizogenes K84 Produces a Hydroxamate Siderophore

Penyalver

Oger

López

et al. 2001

Appl Environ Microbiol

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Iron-binding compounds were produced in various amounts in response to iron starvation by a collection of Agrobacterium strains belonging to the species A. tumefaciens, A. rhizogenes, and A. vitis. The crown gall biocontrol agent A. rhizogenes strain K84 produced a hydroxamate iron chelator in large amounts. Production of this compound, and also of a previously described antibiotic-like substance called ALS84, occurred only in cultures of strain K84 grown in iron-deficient medium. Similarly, sensitivity to ALS84 was expressed only when susceptible cells were tested in low-iron media. Five independent Tn5-induced mutants of strain K84 affected in the production of the hydroxamate iron chelator showed a similar reduction in the production of ALS84. One of these mutants, M8-10, was completely deficient in the production of both agents and grew poorly compared to the wild type under iron-limiting conditions. Thus, the hydroxamate compound has siderophore activity. A 9.1-kb fragment of chromosomal DNA containing the Tn5 insertion from this mutant was cloned and marker exchanged into wild-type strain K84. The homogenote lost the ability to produce the hydroxamate siderophore and also ALS84. A cosmid clone was isolated from a genomic library of strain K84 that restored to strain M8-10 the ability to produce of the siderophore and ALS84, as well as growth in iron-deficient medium. This cosmid clone contained the region in which Tn5 was located in the mutant. Sequence analysis showed that the Tn5 insert in this mutant was located in an open reading frame coding for a protein that has similarity to those of the gramicidin S synthetase repeat superfamily. Some such proteins are required for synthesis of hydroxamate siderophores by other bacteria. Southern analysis revealed that the biosynthetic gene from strain K84 is present only in isolates of A. rhizogenes that produce hydroxamate-type compounds under low-iron conditions. Based on physiological and genetic analyses showing a correlation between production of a hydroxamate siderophore and ALS84 by strain K84, we conclude that the two activities share a biosynthetic route and may be the same compound.Siderophores are relatively low-molecular-weight, iron-chelating agents produced by bacteria and fungi growing under low-iron stress conditions (42). These compounds bind ferric iron (Fe 3ϩ ) with high affinity and transport the metal into the cell via dedicated uptake systems (41). Microbial siderophores vary widely in their overall structure, but most of them contain hydroxamate or catechol groups, which are involved in chelating the ferric ion (42). Apart from their role in the active transport of iron, siderophores may act as growth factors, and some show potent antimicrobial activity (41).The capacity to utilize siderophores is important to the growth of bacteria in the rhizosphere (27) and on plant surfaces (34). Specific siderophore-producing Pseudomonas strains rapidly colonize plant roots of several crops, and this colonization can result in significant yield increas...

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“…Siderophores were produced in M9 medium (31) or Tris minimal salts medium (TMS) (50) supplemented with Casamino Acids (0.3%), tryptophan (0.003%), and thiamine (0.002%). Stock solutions of Casamino Acids were extracted with 8-hydroxyquinoline and chloroform (5) to remove contaminating iron. In some cloning experiments, LB agar was supplemented with 5-bromo-4-chloro-3-indolyl-,-D-galactopyranoside (X-Gal; 40 ,ug/ ml; International Biotechnologies, Inc., New Haven, Conn.) and isopropyl-,B-D-thiogalactopyranoside (IPTG; 100 p,g/ml;…”

Section: Methodsmentioning

confidence: 99%

Cloning and Characterization of Aerobactin Biosynthesis Genes of the Biological Control Agent Enterobacter cloacae

Loper

Ishimaru

Carnegie

et al. 1993

Appl Environ Microbiol

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Five strains of Enterobacter cloacae that are biological control agents of Pythium damping-off diseases produced the hydroxamate siderophore aerobactin under iron-limiting conditions. Genes determining aerobactin biosynthesis of the biocontrol strain E. cloacae EcCT-501 were localized to a 12.3-kb region, which conferred aerobactin production to Escherichia coli DH5a The aerobactin biosynthesis genes of E. cloacae hybridized to those of the pColV-K30 plasmid of E. coli, but restriction patterns of the aerobactin regions of pColV-K30 and E. cloacae differed. A derivative strain with a deletion in the aerobactin biosynthesis locus was as effective as strain EcCT-501 in biological control of Pythium damping-off of cucumber. Thus, aerobactin production did not contribute significantly to the biological control activity of EcCT-501 under the conditions of this study.

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Antibiotic Action of N -Methylthioformohydroxamate Metal Complexes

Cited by 18 publications

References 19 publications

High-affinity iron uptake systems present in Erwinia carotovora subsp. carotovora include the hydroxamate siderophore aerobactin

High-affinity iron uptake systems present in Erwinia carotovora subsp. carotovora include the hydroxamate siderophore aerobactin

Iron-Binding Compounds from Agrobacterium spp.: Biological Control Strain Agrobacterium rhizogenes K84 Produces a Hydroxamate Siderophore

Cloning and Characterization of Aerobactin Biosynthesis Genes of the Biological Control Agent Enterobacter cloacae

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