quinolobactin and pyoverdin

Iron is an essential nutrient for almost all bacteria; however, at neutral pH its bioavailability is limited. Siderophores are iron-binding compounds of low molecular weight that enable the microorganisms that produce them to obtain the necessary iron from the environment. Fluorescent pseudomonads include those that are plant growth promoting, human and plant pathogens, as well as bacteria involved in the biodegradation of xenobiotics. Although pyoverdine is the main siderophore produced by different fluorescent pseudomonads, other siderophores produced by fluorescent pseudomonads include pyochelin, (thio)quinolobactin and pyridine-2, 6-bis thiocarboxylic acid. Research on siderophores continues to reveal new information on their regulation, biosynthesis, function and properties. In this review, we focus on recent advances in the field, particularly on newly characterized siderophores produced by fluorescent pseudomonads and their biotechnological potential.

Topics: Oligopeptides; Proline; Pseudomonas; Quinolines; Siderophores; Thiocarbamates

Under conditions of iron limitation Pseudomonas fluorescens ATCC 17400 produces two siderophores, pyoverdine, and a second siderophore quinolobactin, which itself results from the hydrolysis of the unstable molecule 8-hydroxy-4-methoxy-2-quinoline thiocarboxylic acid (thioquinolobactin). Pseudomonas fluorescens ATCC 17400 also displays a strong in vitro antagonism against the Oomycete Pythium, which is repressed by iron, suggesting the involvement of a siderophore(s). While a pyoverdine-negative mutant retains most of its antagonism, a thioquinolobactin-negative mutant only slowed-down Pythium growth, and a double pyoverdine-, thioquinolobactin-negative mutant, which does not produce any siderophore, totally lost its antagonism against Pythium. The siderophore thioquinolobactin could be purified and identified from spent medium and showed anti-Pythium activity, but it was quickly hydrolysed to quinolobactin, which we showed has no antimicrobial activity. Analysis of antagonism-affected transposon mutants revealed that genes involved in haem biosynthesis and sulfur assimilation are important for the production of thioquinolobactin and the expression of antagonism.

Topics: Antifungal Agents; DNA Transposable Elements; Heme; Hydrolysis; Molecular Sequence Data; Mutation; Oligopeptides; Phenotype; Pseudomonas fluorescens; Pythium; Quinolines; Siderophores; Sulfur

Quinolobactin is a new siderophore produced by a pyoverdine deficient mutant of Pseudomonas fluorescens. A simple and efficient synthesis of quinolobactin is described, starting from xanthurenic acid. The protonation constants of quinolobactin were determined by potentiometric titrations as pKa2 = 5.50+/-0.07, pKa1 = 10.30+/-0.05. The equilibria of the metal complexes were studied by means of spectrophotometric and potentiometric titrations. The overall stability constants of the quinolobactin-FeIII complexes was found to be log beta111 = 18.60+/-0.10, log beta121 = 32.60+/-0.20, log beta120 = 28.20+/-0.25 resulting in a pFeIII value of 18.2 at pH 7.4. The UV-visible spectral parameters of the [FeL2] are in agreement with a complex containing two ligands coordinated to one Fe3+ cation through the oxygen and nitrogen quinoline atoms.

Topics: Gene Expression Regulation, Bacterial; Genes, Bacterial; Hydrogen-Ion Concentration; Iron; Iron Chelating Agents; Iron-Binding Proteins; Kinetics; Ligands; Magnetic Resonance Spectroscopy; Models, Chemical; Nitrogen; Oligopeptides; Oxygen; Potentiometry; Protons; Pseudomonas fluorescens; Quinolines; Siderophores; Spectrophotometry; Ultraviolet Rays

The cytoplasmic membrane protein CcmC is, together with other Ccm proteins, a component for the maturation of c-type cytochromes in Gram-negative bacteria. A Pseudomonas fluorescens ATCC 17400 ccmC mutant is cytochrome c-deficient and shows considerably reduced production of the two siderophores pyoverdine and quinolobactin, paralleled by a general inability to utilize various iron sources, with the exception of haem. The ccmC mutant accumulates in a 5-aminolevulinic acid-dependent synthesis a reddish, fluorescent pigment identified as protoporphyrin IX. As a consequence a visA phenotype similar to that of a ferrochelatase-deficient hemH mutant characterized by drastically reduced growth upon light exposure was observed for the ccmC mutant. The defect of iron-protoporphyrin formation was further demonstrated by the failure of ccmC cell-free proteinase K-treated extracts to stimulate the growth of a haem auxotrophic hemH indicator strain, compared to similarly prepared wild-type extracts. In addition, the ccmC mutant did not sustain hemH growth in cross-feeding experiments while the wild-type did. Significantly reduced resistance to oxidative stress mediated by haem-containing catalases was observed for the ccmC mutant. A double hemH ccmC mutant could not be obtained in the presence of external haem without the hemH gene in trans, indicating that the combination of the two mutations is lethal. It was concluded that CcmC, apart from its known function in cytochrome c biogenesis, plays a role in haem biosynthesis. A function in the regulatory co-ordination of iron acquisition via siderophores, iron insertion into porphyrin via ferrochelatase and iron-protoporphyrin export for cytochrome c formation is predicted.

Topics: Bacterial Proteins; Base Sequence; Biological Transport, Active; Cytochromes c; DNA, Bacterial; Ferrochelatase; Genes, Bacterial; Heme; Iron; Iron Chelating Agents; Membrane Proteins; Models, Biological; Mutation; Oligopeptides; Oxidative Stress; Phenotype; Pigments, Biological; Porphyrins; Protoporphyrins; Pseudomonas fluorescens; Quinolines; Siderophores

Transposon mutant strain 3G6 of Pseudomonas fluorescens ATCC 17400 which was deficient in pyoverdine production, was found to produce another iron-chelating molecule; this molecule was identified as 8-hydroxy-4-methoxy-quinaldic acid (designated quinolobactin). The pyoverdine-deficient mutant produced a supplementary 75-kDa iron-repressed outer membrane protein (IROMP) in addition to the 85-kDa IROMP present in the wild type. The mutant was also characterized by substantially increased uptake of (59)Fe-quinolobactin. The 75-kDa IROMP was produced by the wild type after induction by quinolobactin-containing culture supernatants obtained from the pyoverdine-negative mutant or by purified quinolobactin. Conversely, adding purified wild-type pyoverdine to the growth medium resulted in suppression of the 75-kDa IROMP in the pyoverdine-deficient mutant; however, suppression was not observed when Pseudomonas aeruginosa PAO1 pyoverdine, a siderophore utilized by strain 3G6, was added to the culture. Therefore, we assume that the quinolobactin receptor is the 75-kDa IROMP and that the quinolobactin-mediated iron uptake system is repressed by the cognate pyoverdine.

Topics: Bacterial Outer Membrane Proteins; Culture Media; DNA Transposable Elements; Electrophoresis, Polyacrylamide Gel; Iron; Mutagenesis, Insertional; Oligopeptides; Pigments, Biological; Pseudomonas fluorescens; Quinolines; Siderophores