Johannes C. Liermann

Synthesis of Lamellarin U and Lamellarin G Trimethyl Ether by Alkylation of a Deprotonated α-Aminonitrile

1,2,3,4-Tetrahydroisoquinoline-1-carbonitriles can serve as starting materials for the one-pot synthesis of 5,6-dihydropyrrolo[2,1 a]isoquinolines and 1-benzyl-3,4-dihydroisoquinolines. The latter compounds were transformed to lamellarin G trimethyl ether and lamellarin U in short reaction sequences. This method allows the introduction of acid-sensitive protecting groups for the phenolic hydroxy functions which would be cleaved under the harsh conditions of the classical Bischler-Napieralski reaction.

Sfp-Type 4′-Phosphopantetheinyl Transferase Is Indispensable for Fungal Pathogenicity

In filamentous fungi, Sfp-type 4′-phosphopantetheinyl transferases (PPTases) activate enzymes involved in primary (α-aminoadipate reductase [AAR]) and secondary (polyketide synthases and nonribosomal peptide synthetases) metabolism. We cloned the PPTase gene PPT1 of the maize anthracnose fungus Colletotrichum graminicola and generated PPTase-deficient mutants (Δppt1). Δppt1 strains were auxotrophic for Lys, unable to synthesize siderophores, hypersensitive to reactive oxygen species, and unable to synthesize polyketides (PKs). A differential analysis of secondary metabolites produced by wild-type and Δppt1 strains led to the identification of six novel PKs. Infection-related morphogenesis was affected in Δppt1 strains. Rarely formed appressoria of Δppt1 strains were nonmelanized and ruptured on intact plant. The hyphae of Δppt1 strains colonized wounded maize (Zea mays) leaves but failed to generate necrotic anthracnose disease symptoms and were defective in asexual sporulation. To analyze the pleiotropic pathogenicity phenotype, we generated AAR-deficient mutants (Δaar1) and employed a melanin-deficient mutant (M1.502). Results indicated that PPT1 activates enzymes required at defined stages of infection. Melanization is required for cell wall rigidity and appressorium function, and Lys supplied by the AAR1 pathway is essential for necrotrophic development. As PPTase-deficient mutants of Magnaporthe oryzea were also nonpathogenic, we conclude that PPTases represent a novel fungal pathogenicity factor.

Hirsutane-Type Sesquiterpenes with Uncommon Modifications from Three Basidiomycetes

From three basidiomycetes, Xeromphalina sp., Stereum sp., and Pleurocybella porrigens, six triquinane sesquiterpenes with unprecendented modifications and a rearranged sesquiterpene related to coriolin C have been isolated. Their isolation, structure elucidation, and biological evaluation are described.

Xanthepinone, an antimicrobial polyketide from a soil fungus closely related to Phoma medicaginis.

The isolation, biological characterization, and structure elucidation of xanthepinone, a novel antifungal metabolite isolated from the broth of submerged cultures of a soil fungus, are described. Xanthepinone inhibits the conidial germination of Magnaporthe grisea (2 microg/mL), Phytophthora infestans (5 microg/mL), and Botrytis cinerea (10 microg/mL) while showing only weak antibacterial activity; cytotoxicity was not observed up to 50 microg/mL. Molecular taxonomy revealed that the producing strain is close to species in the genus Phoma as well as to uncultured soil fungi and endophytes.

Oxacyclododecindione, a novel inhibitor of IL-4 signaling from Exserohilum rostratum.

In a screening program for new metabolites from fungi inhibiting the IL-4 mediated signal transduction, a novel chlorinated macrocyclic lactone, designated as oxacyclododecindione, was isolated from fermentations of the imperfect fungus Exserohilum rostratum. The structure was determined by a combination of spectroscopic techniques. Oxacyclododecindione inhibits the IL-4 induced expression of the reporter gene secreted alkaline phosphatase (SEAP) in transiently transfected HepG2 cells with IC50 values of 20~25 ng/ml (54~67.5 nM). Studies on the mode of action of the compound revealed that the inhibition of the IL-4 dependent signaling pathway is caused by blocking the binding of the activated STAT6 transcription factors to the DNA binding site without inhibiting tyrosine phosphorylation. The compound has no antibacterial or antifungal activity.

Tetracyclic Terpenoids from Dasyscyphus niveus, Dasyscyphins D and E

Cultures of the ascomycete Dasyscyphus niveus have yielded two new tetracyclic dasyscyphin-type terpenoids (1 and 2), and their structures were elucidated by NMR spectroscopy and X-ray crystallography. The absolute configuration of dasyscyphin D (1) was determined by synthesis and NMR spectroscopy of diastereomeric MTPA esters. Both compounds inhibited the germination of conidia of Magnaporthe grisea at 25 microg/mL.

Phytotoxic dioxolanone-type secondary metabolites from Guignardia bidwellii.

Phenguignardic acid was recently described as a phytotoxic secondary metabolite from submerged cultures of the grape black rot fungus Guignardia bidwellii. Since the production rate of this natural product in submerged culture is very low, fermentation optimisation was carried out. The optimisation of cultivation conditions led to the identification of seven secondary metabolites, structurally related to guignardic acid, a known secondary metabolite from Guignardia species containing a dioxolanone moiety. All metabolites presented here have not been described to date and are presumably biosynthesised via deamination products of amino acids, such as phenylalanine, valine, tyrosine, and alanine. Four of the seven compounds showed phytotoxic activity. Based on the structures determined by NMR spectroscopy a preliminary structure activity relationship indicated a free carboxyl group as presumably required for the phytotoxic activity.

Phenguignardic acid and guignardic acid, phytotoxic secondary metabolites from Guignardia bidwellii.

Bioactivity-guided isolation led to the identification of phenguignardic acid (2), a new phytotoxic secondary metabolite from submerged cultures of grape black rot fungus, Guignardia bidwellii. The compound is structurally related to guignardic acid (1), a dioxolanone moiety-containing metabolite isolated previously from Guignardia species. However, in contrast to guignardic acid, which is presumably synthesized from deamination products of valine and phenylalanine, the biochemical precursor for the biosynthesis of the new phytotoxin appears to be exclusively phenylalanine. Guignardic acid was also found in extracts of cultures from Guignardia bidwellii. The phytotoxic activities of both compounds were assessed in plant assays using either detached vine leaves or intact plants. Antimicrobial and cytotoxic activities of phenguignardic acid were determined.

Allantofuranone, a new antifungal antibiotic from Allantophomopsis lycopodina IBWF58B-05A

In a screening for new bioactive compounds, the extract of Allantophomopsis lycopodina strain IBWF58B-05A, an imperfect ascomycete, was found to exhibit strong but rather selective antibiotic activity against Paecilomyces variotii. The bioactivity-guided isolation yielded allantofuranone, a new and uncommon γ-lactone. This compound showed antifungal activity against P. variotii and Penicillium species. This paper describes the isolation, structure elucidation and biological characteristics of allantofuranone.

Caripyrin, a new inhibitor of infection-related morphogenesis in the rice blast fungus Magnaporthe oryzae.

Caripyrin (trans-5-(3-methyloxiranyl)pyridincarboxylic acid methyl ester, 1), a new pyridyloxirane, was isolated from submerged cultures of the basidiomycete Caripia montagnei. The compound was found to inhibit conidial germination and appressorium formation in the rice blast fungus Magnaporthe oryzae, whereas the infection-related morphogenesis in several other phytopathogenic fungi was not affected. In plant assays on rice, 1 was found to protect plants more efficiently against fungal infection than the structurally related fungal secondary metabolite, fusaric acid. Contrary to the latter, 1 was neither cytotoxic, antibacterial, nor nematicidal.