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G Winkelmann

Publications and source records attributed to G Winkelmann.

At least 37 records · Page 2Linked to original sources
PubMed1998

Molecular recognition of siderophores: a study with cloned ferrioxamine receptors (FoxA) from Erwinia herbicola and Yersinia enterocolitica.

The outer membrane receptor for ferrioxamines (FoxAErw) of Erwinia herbicola (Pantoea agglomerans) was cloned from a cosmid gene bank and partially sequenced. A comparison of the partial amino acid sequence of FoxAErw with the amino acid sequence of FoxAYer from Yersinia enterocolitica revealed a high sequence homology. A functional analysis of FoxAErw and FoxAYer receptors cloned into a Fhu-negative background (HK97) revealed that ferrioxamines are recognized at very low concentrations (< 10 pmoles) in growth promotion bioassays. A collection of ferrioxamine derivatives containing varying chain lengths and ether bridges within the molecule was also accepted. However, the three ether containing ferrioxamine (Et3) behaved differently in the two FoxA receptors. Coprogen was also recognized to a certain extent, whereas ferrichromes were completely excluded from the FoxA receptors, confirming that coprogens share some structural similarities with the ferrioxamines. FoxA mutants (FM13) of Erwinia herbicola obtained by ferrimycin selection showed no uptake of 55Fe-labelled ferrioxamine E and B any more, while the transport of coprogen and ferrichrome was unaffected or even slightly increased.

Amino Acid Sequence↗
PubMed1998

Pyrrolnitrin from Burkholderia cepacia: antibiotic activity against fungi and novel activities against streptomycetes.

A bacterial strain identified as Burkholderia cepacia NB-1 was isolated from water ponds in the botanical garden in Tübingen, Germany, and was found to produce a broad spectrum phenylpyrrole antimicrobial substance active against filamentous fungi, yeasts and Gram-positive bacteria. In batch culture containing glycerol and L-glutamic acid, the isolate NB-1 produced the antibiotic optimally late in the growth phase and accumulated a main portion in their cells. Isolation and purification of the antibiotic from Burkholderia (Pseudomonas) cepacia NB-1 by acetone extraction, gel filtration on Sephadex LH-20 and preparative HPLC yielded 0.54 mg l-1 of a pure substance. Spectroscopic data (HPLC, MS and NMR) confirmed that the compound was pyrrolnitrin [3-chloro-4-(2'-nitro-3'-chloro-phenyl) pyrrole]. Pyrrolnitrin has an inhibitory effect on the electron transport system, as demonstrated by isolated mitochondria from Neurospora crassa 74 A. This inhibition was relieved by N,N,N',N'-tetramethyl-p-phenylenediamine dihydrochloride (TMPD), indicating that pyrrolnitrin blocked the electron transfer between the dehydrogenases and the cytochrome components of the respiratory chain. Among Gram-positive bacteria, pyrrolnitrin was most active against certain Streptomyces species, especially S. antibioticus, which has not previously been described in the literature. In the presence of pyrrolnitrin, aerial mycelium and spore formation of Strep. antibioticus was suppressed, although growth continued via substrate mycelium. The new findings of inhibition of streptomycetes and their secondary metabolism by pyrrolnitrin may contribute to the fact that Pseudomonas species predominate in soil and compete even with antibiotic-producing Streptomyces.

Antifungal Agents↗
PubMed1998

Combination of behaviour therapy with fluvoxamine in comparison with behaviour therapy and placebo. Results of a multicentre study.

BACKGROUND: We investigated whether the combination of multi-modal behaviour therapy (BT) with fluvoxamine is superior to BT and placebo in the acute treatment of severely ill in-patients with obsessive-compulsive disorder (OCD). METHOD: In a randomised, double-blind design, 30 patients were treated for nine weeks with BT plus placebo and 30 patients with BT plus fluvoxamine (maximum dosage 300 mg, mean dose 288.1 mg). BT included exposure with response prevention, cognitive restructuring and development of alternative behaviours. RESULTS: Both groups showed a highly significant symptom reduction after treatment. There were no significant differences between the groups concerning compulsions. Obsessions were significantly more reduced in the fluvoxamine and BT group than in the placebo and BT group. Furthermore, the group BT plus fluvoxamine showed a significantly higher response rate (87.5 v. 60%) according to a previously defined response criterion. Severely depressed patients with OCD receiving BT plus placebo presented a significantly worse treatment outcome (Y-BOCS scores) than all other groups. CONCLUSIONS: The results suggest that BT should be combined with fluvoxamine when obsessions dominate the clinical picture and when a secondary depression is present.

Adult↗
PubMed1997

Drug specific antibodies: T-cell epitope-lipopeptide conjugates are potent adjuvants for small antigens in vivo and in vitro.

To generate conventional or monoclonal antibodies for the serological detection of drugs, antibiotics, toxins and other low molecular mass substances, a suitable and effective adjuvant is needed. Lipopeptides derived from a major component of the bacterial cell wall constitute potent nontoxic and nonpyrogenic immunoadjuvants when mixed with conventional antigens. Here we demonstrate that the synthetic lipopeptide N-palmitoyl-S-[2,3-bis(palmitoyloxy)-(2R,S)-propyl]-(R)-cysteinyl- serine (P3CS) coupled to a Th-cell epitope (P3CS-Th) can efficiently enhance the specific immune response against low molecular weight compounds in different species. In the presence of the synthetic lipopeptide P3CS-Th, the peptides which are per se non-immunogenic stimulated a specific humoral immune response in mice after intraperitoneal application. Mixtures containing adjuvants without the Th sequence showed no significant antibody induction. A marked enhancement of the humoral immune response was obtained with the low molecular mass antigens Iturin AL, Herbicolin A and Microcystin (MLR) coupled to poly-l-lysin (MLR-PLL), in rabbits and in chickens. Lipopeptide-Th cell epitope conjugates also constituted adjuvants for the in vitro immunization of either human mononuclear cells or mouse B-cells with MLR-PLL; after fusion of the immunized cultures with the heteromyeloma cell lines CB-F7 or the mouse myeloma cell line SP 2/0, respectively, we observed a significantly increased yield of antibody secreting hybridomas.

Adjuvants, Immunologic↗
PubMed1996

Characterization of a novel Spirillum-like bacterium that degrades ferrioxamine-type siderophores.

A novel Gram-negative Spirillum-like bacterium (ASP-1) was isolated from lake water by enrichment culture on desferrioxamine B as sole source of carbon and energy. ASP-1 was able to degrade the siderophores desferrioxamine B and E. The property of siderophore degradation was inducible in the presence of desferrioxamine B. The ferric complexes, however, were not measurably degraded but served as an iron source. Degradation of desferrioxamines in culture was followed by measuring the residual ferrioxamines colorimetrically at 430 nm after addition of iron. Degradation in cell-free assays was followed quantitatively by HPLC on a reversed-phase column measuring the time-dependent disappearance of the desferrioxamines B and E. Cell-free assays also revealed that degradation of the cyclic desferrioxamine E was rapid and complete, whereas degradation of the linear desferrioxamine B yielded two intermediate iron-binding metabolites of shorter chain length. Preparative isolation by HPLC and mass spectrometric analysis of the metabolites revealed masses at 361 and 419 a.m.u., respectively, suggesting a splitting at the two amide bonds. ASP-1 is a nitrogen fixing Spirillum bacterium which could also use ammonium and glucose or several organic acids as a carbon source but grew poorly with amino acids. Physiological comparisons with Aquaspirillum and Azospirillum failed to assign ASP-1 to any of the presently known Spirillum species. Based on 16S rDNA sequence analysis the strain could be placed within the radiation of the Azospirillum/Rhodocista group. The closest relative was Azospirillum irakense, showing 98.8% similarity.

Biodegradation, Environmental↗
PubMed1996

Maltophilin: a new antifungal compound produced by Stenotrophomonas maltophilia R3089.

Strains of Stenotrophomonas maltophilia R3089, isolated from the rhizosphere of rape plants (Brassica napus L.), produced a novel antifungal compound, named maltophilin, which inhibited the growth of various saprophytic, human-pathogenic and phytopathogenic fungi but was inactive against Gram-positive and Gram-negative bacteria. Maltophilin is a novel macrocyclic lactam antibiotic with a molecular mass of 510 mu. The compound was isolated from the culture filtrate by ethyl acetate extraction and gel filtration on Sephadex LH20 and purified by preparative HPLC on reversed phase. The structure of maltophilin was elucidated by electrospray mass spectrometry and 1H and 13C NMR spectroscopy.

Anti-Bacterial Agents↗
PubMed1995

Ornibactin production and transport properties in strains of Burkholderia vietnamiensis and Burkholderia cepacia (formerly Pseudomonas cepacia).

Several strains of Burkholderia vietnamiensis, isolated from the rhizosphere of rice plants, and four strains formerly known as Pseudomonas cepacia including two collection strains and two clinical isolates were compared for siderophore production and iron uptake. The B. vietnamiensis (TVV strains) as well as the B. cepacia strains (ATCC 25416 and ATCC 17759) and the clinical isolates K132 and LMG 6999 were all found to produce ornibactins under iron starvation. The two ATCC strains of B. cepacia additionally produced the previously described siderophores, pyochelin and cepabactin. Analysis of the ratio of isolated ornibactins (C4, C6 and C8) by HPLC revealed nearly identical profiles. Supplementation of the production medium with ornithine (20 mM) resulted in a 2.5-fold increase in ornibactin synthesis. Ornibactin-mediated iron uptake was independent of the length of the acyl side chain and was observed with all strains of B. vietnamiensis and B. cepacia, but was absent with strains of Pseudomonas aeruginosa, Pseudomonas fluorescens and Pseudomonas stutzeri, known to produce pyoverdines or desferriferrioxamines as siderophores. These results suggest that ornibactin production is a common feature of all Burkholderia strains and that these strains develop an ornibactin-specific iron transport system which is distinct from the pyoverdine-specific transport in Pseudomonas strains.

Biological Transport↗
PubMed1995

A seizure, and electroencephalographic signs of a lowered seizure threshold, associated with fluvoxamine treatment of obsessive-compulsive disorder.

A 38-year-old patient with severe obsessive-compulsive disorder received fluvoxamine in a clinical study. Psychometric ratings showed marked clinical improvement in the third week of fluvoxamine administration, but after 8 weeks, at a dose of 300 mg per day, he suffered a grand mal seizure after drinking a glass of beer (0.2 liter). He had no history of previous epileptic seizures. Careful neurological evaluation including computer tomography and magnetic resonance imaging of the brain revealed no signs of acute disease. EEG before the fit did not show epileptiform activity; after the fit, spikes and spike-wave complexes appeared, which disappeared upon discontinuation of fluvoxamine. Since his obsessive-compulsive symptoms had responded well to fluvoxamine and worsened after its discontinuation, the drug was cautiously reintroduced. Improvement of the obsessive-compulsive symptoms was observed again, but spikes and spike-wave complexes reappeared at a dose of 50 mg per day. Under anticonvulsant treatment with carbamazepine, fluvoxamine was increased to 100 mg per day. No seizures occurred during a follow-up to two years.

Adult↗
PubMed1994

Iron supply of Pasteurella multocida and Pasteurella haemolytica.

Pasteurella multocida and Pasteurella haemolytica do not produce hydroxamate- or phenolate type siderophores. However, transport- and utilization systems could be detected for the well known siderophores ferrioxamine B, E, G, rhizoferrin and the intermediate 2,3-dihydroxybenzoic acid by means of cross-feeding tests in both Pasteurella species. Enterobactin and ferrichrome did not feed any of the Pasteurella strains tested. Additionally, alpha-ketoacids and alpha-hydroxyacids such as pyruvic acid, alpha-hydroxyisovaleric acid and others acting as primary metabolites enabled growth of P. multocida and P. haemolytica under iron limitation.

Culture Media↗
PubMed1994

HPLC separation of enterobactin and linear 2,3-dihydroxybenzoylserine derivatives: a study on mutants of Escherichia coli defective in regulation (fur), esterase (fes) and transport (fepA).

Reversed-phase HPLC separation of enterobactin and its 2,3-dihydroxybenzoylserine derivatives was used for a comparative analysis of mutants of Escherichia coli, defective in the regulation of enterobactin biosynthesis (fur), enterobactin transport (fepA) and enterobactin esterase (fes). A complete separation of all 2,3-dihydroxybenzoylserine compounds was achieved: the monomer (DHBS), the linear dimer (DHBS)2 and trimer (DHBS)3, the cyclic trimer, enterobactin, as well as 2,3-dihydroxybenzoic acid. The production of all these compounds was followed after ethylacetate extraction from acidified culture fluids. Enterobactin was found to be the predominant product in all mutant strains. The mutant strains behaved differently with regard to the breakdown products. All degradation products, such as DHBS, (DHBS)2 and (DHBS)3, were detected in the overproducing fur mutant where both transport and esterase are still functioning, while only the monomer, DHBS, was detected in the fepA mutant and no degradation was found in the esterase-deficient fes mutant. From the pattern of breakdown products it may be inferred that the esterase acts in two different ways, depending on whether transport is functioning or not. Thus, esterolytic cleavage of ferric enterobactin after entering the cells results in a mixture of all three hydrolysis products, i.e. DHBS, (DHBS)2 and (DHBS)3, while cleavage of iron-free enterobactin subsequent to its biosynthesis yields only the monomer. Thus, the results of quantitative HPLC analysis of enterobactin and its breakdown products show that different enterobactin esterase products arise, depending on whether iron is bound to enterobactin or not.

Bacterial Outer Membrane Proteins↗
PubMed1994

Sleep EEG of patients with obsessive-compulsive disorder.

Twenty-two patients suffering from an obsessive and compulsive disorder (OCD) according to DSM-III-R were investigated by polysomnographic sleep EEG recordings under drug-free conditions and compared to age- and sex-matched healthy controls. Sleep efficiency was significantly lower and wake % SPT was significantly increased in the patient group compared to healthy subjects. Sleep architecture did not differ among the two samples. Especially REM sleep measures, in particular, REM latency did not differ among the groups. No positive correlation was found between sleep variables and rating inventories for obsession and compulsions (Y-BOCS), depression (Hamilton) and anxiety (CAS). A secondary depression did not influence sleep EEG variables. The results of this study contradict the assumption that OCD patients show REM sleep and slow wave sleep abnormalities similar to those shown by patients with primary depression.

Adult↗
PubMed1993

Ornibactins--a new family of siderophores from Pseudomonas.

Novel linear hydroxamate/hydroxycarboxylate siderophores from strains of Pseudomonas cepacia were isolated and named ornibactins. The ornibactins represent modified tetrapeptide siderophores, possessing the sequence L-Orn1(N delta-OH, N delta-acyl)-D-threo-Asp(beta-OH)-L-Ser-L-Orn4(N delta-OH, N delta-formyl)-1,4-diaminobutane. The N delta-acyl groups of Orn1(N delta-OH, N delta-acyl) may vary and represent the three acids 3-hydroxybutanoic acid, 3-hydroxyhexanoic acid and 3-hydroxyoctanoic acid, leading to a mixture of three different ornibactins, designated according to their acyl chain length as ornibactin-C4, ornibactin-C6 and ornibactin-C8. Each of the siderophores is accompanied by a small amount of a more hydrophilic component with a 16 a.m.u. higher mass. The structure elucidation was based on results from gas chromatography amino acid analysis, electrospray mass spectrometry, and one- and two-dimensional nuclear magnetic resonance techniques.

Amino Acid Sequence↗
PubMed1993

Alpha-keto acids are novel siderophores in the genera Proteus, Providencia, and Morganella and are produced by amino acid deaminases.

Growth promotion and iron transport studies revealed that certain alpha-keto acids generated by amino acid deaminases, by enterobacteria of the Proteus-Providencia-Morganella group (of the tribe Proteeae), show significant siderophore activity. Their iron-binding properties were confirmed by the chrome azurol S assay and UV spectra. These compounds form ligand-to-metal charge transfer bands in the range of 400 to 500 nm. Additional absorption bands of the enolized ligands at 500 to 700 nm are responsible for color formation. Siderophore activity was most pronounced with alpha-keto acids possessing an aromatic or heteroaromatic side chain, like phenylpyruvic acid and indolylpyruvic acid, resulting from deamination of phenylalanine and tryptophan, respectively. In addition, alpha-keto acids possessing longer nonpolar side chains, like alpha-ketoisocaproic acid or alpha-ketoisovaleric acid and even alpha-ketoadipic acid, also showed siderophore activity which was absent or negligible with smaller alpha-keto acids or those possessing polar functional groups, like pyruvic acid, alpha-ketobutyric acid, or alpha-ketoglutaric acid. The fact that deaminase-negative enterobacteria, like Escherichia coli and Salmonella spp., could not utilize alpha-keto acids supports the view that specific iron-carboxylate transport systems have evolved in members of the tribe Proteeae and are designed to recognize ferric complexes of both alpha-hydroxy acids and alpha-keto acids, of which the latter can easily be generated by L-amino acid deaminases in an amino acid-rich medium. Exogenous siderophores, like ferric hydroxamates (ferrichromes) and ferric polycarboxylates (rhizoferrin and citrate), were also utilized by members of the tribe Proteeae.

Amino Acids↗
PubMed1992

Rhizoferrin: a complexone type siderophore of the Mucorales and entomophthorales (Zygomycetes).

The present investigation presents evidence that rhizoferrin, a novel polycarboxylate or complexone-type siderophore, originally isolated from Rhizopus microsporus, represents the common siderophore within the Zygomycetes. Thus, rhizoferrin could be detected by HPLC analysis in various families of the Mucorales, e.g., Rhizopus microsporus var. rhizopodiformis, Mucor mucedo and Phycomyces nitens (Mucoraceae), Chaetostylum fresenii and Cokeromyces recurvatus (Thamnidiaceae), Cunninghamella elegans and Mycotypha africana (Choanephoraceae) and Mortierella vinacea (Mortierellaceae) and in Basidiobolus microsporus (Entomophthorales). The function of rhizoferrin as a siderophore in the fungus R. microsporus var. rhizopodiformis was demonstrated by time- and concentration-dependent uptake of [55Fe]-labelled rhizoferrin, yielding saturation kinetics with values of Km = 8 microM and V(max) = 1.2 nmol min-1 (mg dry wt)-1.

Fungi↗
PubMed1991

Fermentation and isolation of herbicolin A, a peptide antibiotic produced by Erwinia herbicola strain A 111.

Erwinia herbicola (Enterobacter agglomerans), belonging to the Enterobacteriaceae, produces the lipopeptide antibiotics herbicolin A and B, which are active against sterol-containing fungi. Fermentation of these antibiotics was performed in 20-l stirred glass fermentors in a batch process. Best yields of antibiotic production were found at low cultivation temperatures in a TRIS-buffered chemically defined medium. Under these conditions the amount of impurities aggravating the purification was minimized. Isolation was performed by adsorption, and gel and ion exchange chromatographic techniques. In a final purification step preparative high performance liquid chromatography (PHPLC) yielded pure herbicolin A.

Amino Acid Sequence↗
PubMed1991

Identification of enterobactin and linear dihydroxybenzoylserine compounds by HPLC and ion spray mass spectrometry (LC/MS and MS/MS).

The production of catecholate siderophores was studied in some selected species of Enterobacter (Enterobacteriaceae). The extracted catecholates were separated as iron-free compounds by HPLC on a C18 reversed-phase column using methanol/0.1% phosphoric acid or methanol/0.1% formic acid as a solvent system and identified by ion spray mass spectrometry (LC/MS, MS/MS). Five catecholate compounds were identified which include 2,3-dihydroxybenzoylserine, its linear dimer and trimer, the cyclic enterobactin and an unidentified isomer of enterobactin. In addition, a new large-scale method for the isolation of catecholate siderphores is described which is based on adsorption on XAD-2 and subsequent purification on Sephadex LH20.

Biopolymers↗