Structure, production characteristics and fungal antagonism of tensin - a new antifungal cyclic lipopeptide from Pseudomonas fluorescens strain 96.578.

AIM: To study the antagonistic activity by Pseudomonas fluorescens strain 96.578 on the plant pathogenic fungus Rhizoctonia solani. METHODS AND RESULTS: Strain 96.578 produced a new cyclic lipopeptide, tensin. High tensin production per cell was detected in liquid media with glucose, mannitol or glutamate as growth substrate while fructose, sucrose and asparagine supported low production. Tensin production was nearly constant in media with different initial C levels, while low initial N contents reduced production. When applied to sugar beet seeds, strain 96.578 produced tensin during seed germination. When challenged with strain 96.578 or purified tensin, Rhizoctonia solani reduced radial mycelium extension but increased branching and rosette formation. CONCLUSION: The antagonistic activity of strain 96.578 towards Rhizoctonia solani was caused by tensin. SIGNIFICANCE AND IMPACT OF THE STUDY: When coated onto sugar beet seeds, tensin production by strain 96.578 could be of significant importance for inhibition of mycelial growth and seed infection by Rhizoctonia solani.

Viscosinamide, a new cyclic depsipeptide with surfactant and antifungal properties produced by Pseudomonas fluorescens DR54.

Pseudomonas fluorescens DR54 showed antagonistic properties against plant pathogenic Pythium ultimum and Rhizoctonia solani both in vitro and in planta. Antifungal activity was extractable from spent growth media, and fractionation by semi-preparative HPLC resulted in isolation of an active compound, which was identified as a new bacterial cyclic lipodepsipeptide, viscosinamide, using 1D and 2D 1H-, 13C-NMR and mass spectrometry. The new antibiotic has biosurfactant properties but differs from the known biosurfactant, viscosin, by containing glutamine rather than glutamate at the amino acid position 2 (AA2). No viscosin production was observed, however, when Ps. fluorescens DR54 was cultured in media enriched with glutamate. In vitro tests showed that purified viscosinamide also reduced fungal growth and aerial mycelium development of both P. ultimum and R. solani. Viscosinamide production by Ps. fluorescens DR54 was tightly coupled to cell proliferation in the batch cultures, as the viscosinamide produced per cell mass unit approached a constant value. In batch cultures with variable initial C, N or P nutrient levels, there were no indications of elevated viscosinamide production during starvation or maintenance of the cultures in stationary phase. Analysis of cellular fractions and spent growth media showed that a major fraction of the viscosinamide produced remained bound to the cell membrane of Ps. fluorescens DR54. The isolation, determination of structure and production characteristics of the new compound with both biosurfactant and antibiotic properties have promising perspectives for the application of Ps. fluorescens DR54 in biological control.

Reactions of N-N and N-O compounds with horseradish peroxidase and peroxidases from Mycobacterium tuberculosis.

Several N-N-and N-O-containing compounds were analysed for their ability to act as substrates for horseradish peroxidase and peroxidases in Mycobacterium tuberculosis extracts. Aminoguanidine, diaminoguanidine, isoniazid, hydroxylamine and hydrazine were found to be weak substrates for horseradish peroxidase in reaction I and to inhibit the reaction of horseradish peroxidase with hydrogen peroxide. The same compounds inhibited the reaction of Mycobacterium tuberculosis peroxidase-catalase with hydrogen peroxide, and hydroxylamine was found to be a weak substrate for this enzyme. In growth inhibition experiments, diaminoguanidine inhibited the growth of M. tuberculosis H37Rv at 50 microg/mL, but not the growth of two isoniazid-resistant strains. Isonicotinic acid hydroxamate inhibited the reaction of the peroxidases with hydrogen peroxide, but was not itself a substrate and had no growth-inhibitory effects. On the basis of these results we suggest that the effect of isoniazid on growth of M. tuberculosis results from increased oxidative stress due to inhibition of catalase-peroxidase as well as from generation of toxic radicals with the structure [structure in text].

N-Acyl-L-homoserine lactone autoinducers control production of an extracellular lipopeptide biosurfactant required for swarming motility of Serratia liquefaciens MG1.

A nonswarming Serratia liquefaciens mutant deficient in serrawettin W2 production was constructed by transposon mutagenesis. Sequence homology indicated that insertion had occurred in gene swrA, which encodes a putative peptide synthetase. Expression of swrA is controlled by quorum sensing.

Triterpenoid saponins from Phytolacca rivinoides and Phytolacca bogotensis.

Investigation of the ethanolic extracts from Phytolacca rivinoides and P. bogotensis has resulted in the isolation of five new triterpenoid glycosides of serjanic acid. Their structures have been established mainly by spectroscopic methods (FAB-MS, 1H, 13C NMR, COSY, NOESY, TOCSY, HETCOR and J-resolved 1H NMR) as 3-O-(O-beta-D-galactopyranosyl-(1-->3)-O-beta-D-glucopyranosyl)serjan ic acid 28-O-beta-D-glucopyranosyl ester, 3-O-(O-beta-D-glucopyranosyl-(1-->3)-O-[beta-D-galactopyranosyl-(1-->4)] -O- beta-D-glucopyranosyl)serjanic acid 28-O-beta-D-glucopyranosyl ester, 3-O-(O-alpha-L-rhamnopyranosyl-(1-->2)-O-beta-D-glucopyranosyl-(1-->2)- O-beta-glucopyranosyl)serjanic acid 28-O-beta-D-glucopyranosyl ester, 3-O-(O-beta-D-glucopyranosyl-(1-->3)-O-beta-D-galactopyranosyl-(1-->4)- O-beta-D-glucopyranosyl)serjanic acid 28-O-beta-D-glucopyranosyl ester and 3-O-(O-beta-D-galactopyranosyl-(1-->4)-O-[beta-D-glucopyranosyl-(1-->3)] - O-beta-D-glucopyranosyl)serjanic acid.

Two new flavones from Calliandra californica.

Two new flavones were isolated from the medicinal plant Calliandra californica, 7,2',4',5'-tetramethoxyflavone [1] and 5-hydroxy-7,2',4',5'-tetramethoxyflavone [2]. The structures were determined from spectral evidence and both compounds were synthesized. Compound 2 exhibited antimicrobial activity for two test bacterial strains.

Marine alkaloids, 15. Two alkaloids, flustramine E and debromoflustramine B, from the marine bryozoan Flustra foliacea.

A new alkaloid, flustramine E [1], with inhibitory activity towards Rhizotonia solani and Botrytis cinerea was isolated by gas-phase extraction from the marine bryozoan Flustra foliacea. The structure was determined as 3a,8a-cis-1-methyl-3a-(3-methyl-2- butenyl)-6-bromo-1,2,3,3a,8,8a-hexahydropyrrolo[2,3-b]indole, based on spectroscopic investigations. Another alkaloid, debromoflustramine B [2a], was detected in trace amounts and identified by comparison of the mass spectrum with that of a synthetic sample. The extract also contained the previously reported alkaloids flustramine A and B in the ratio 1:7.

Albaflavenone, a sesquiterpene ketone with a zizaene skeleton produced by a streptomycete with a new rope morphology.

A novel antibiotic alpha,beta-unsaturated sesquiterpene ketone, albaflavenone with a zizaene skeleton was isolated from a morphologically novel, highly odorous Streptomyces species which was identified with the species group S. albidoflavus, cluster 1. The new compound, partly responsible for the odour, was assigned the structure of 2R',6,7,7-tetramethyl-1S',8R'- tricyclo-[6.2.1.0(1,5)]undec-5-en-4-one based on spectroscopic studies including 2D NMR (COSY, HETCOR, ROESY, NOE-difference) experiments.

Poisonings from flesh of the Greenland shark Somniosus microcephalus may be due to trimethylamine.

The flesh of the Greenland shark, Somniosus microcephalus, especially in fresh condition, is toxic to both dog and man. Analysis of the toxic fractions indicates the presence of large amounts of trimethylamine oxide (TMAO) but no other substances that could be directly responsible for the poisoning. The symptoms appear to be due to an acute trimethylamine (TMA) poisoning arising from intestinal reduction of TMAO to TMA. TMA stimulates contraction of the guinea-pig ileum (antagonized by atropine but not hexamethonium). Low concentrations of TMA increase contraction of the rat phrenic nerve-diaphragm preparation, whereas higher concentrations cause blockade.

Tetramine: occurrence in marine organisms and pharmacology.

The occurrence of tetramethylammonium ion (TMA) in marine organisms is reviewed. The pharmacological action of TMA is also discussed, with special emphasis on the sign and symptoms experienced by oral poisonings. It is concluded that the major manifestations of TMA poisonings may be attributed to interactions with the autonomous nervous system. Fatal intoxications are due to neuromuscular blockade.

The toxin tetramine from the "edible" whelk Neptunea antiqua.

The chemical nature of extracts of the marine gastropod Neptunea antiqua collected in the North Sea has been investigated. Amino acid analysis and spectroscopical studies (FAB MS, 1H NMR, 13C NMR and IR) on fractions purified by Biogel P-2 and Amberlite IR 120 column chromatography allowed the identification of 19 components. It was established that the water-soluble toxin responsible for poisonings, following ingestion of this snail, is tetramine (the tetramethylammonium ion present as an unknown salt). Contrary to what occurs in other Neptunea species (N. arthritica and N. intersculpta), tetramine was found not only in the salivary gland, but also in the remaining part of the animal, albeit in smaller concentration. In the isolated guinea-pig ileum assay, synergistic effects with other main components present (betaine, homarine) could not be demonstrated. Choline esters, believed to act synergistically in other Neptunea species, were not detected in Neptunea antiqua.

Diagnostic positive and negative ion fast atom bombardment mass spectrometry of low-molecular-weight ammonium compounds of marine origin.

Five ammonium compounds--betaine, taurine, homarine, trigonelline and tetramethylammonium chloride--were investigated by positive and negative ion fast atom bombardment (FAB) mass spectrometry. Observation of the parent cation [M + H]+ together with series of clusters (e.g. [nM + H]+, n = 2-5) and fragmentation patterns in the positive FAB mode may be used diagnostically as a fingerprint of the compound. The shifts in mass and relative intensity by addition of trichloroacetic acid and metal salts offer corroborating information of the identity of a compound in a mixture. The FAB negative ion spectra were characterized by clusters incorporating the anion (e.g. [M + Cl]-) but are generally less informative than the spectra obtained in the positive mode.