No abstract

From the culture broth of the mould Trichoderma viride, strain NRRL 3199, a microheterogeneous mixture of the membrane active 20-residue peptaibol alamethicin (ALM) could be isolated. ALMs were isolated by XAD-2 column chromatography and separated by silica gel chromatography and trichloromethane/MeOH gradient elution into an acidic and neutral group of peptides, named ALM F30 and ALM F50, respectively, according to their 100 Rf on TLC. Peptides ALM F50 were separated by semi-preparative and analytical HPLC and subjected to ESI-MS. Ten sequences of ALM F30 and their relative quantities could be determined. The major peptides ALM F30/3 (46%) and ALM F30/7 (40%), distinguished by Aib/Ala exchange in position 6, correspond to sequences described as ALM I and II occurring in the original alamethicin from Upjohn Company. Analogously, 13 sequences of the neutral peptide mixture named ALM F50 could be determined. The major peptide ALM F50/5 (75%) and the minor peptide ALM F50/7 (10%) are distinguished from ALM F30/3 and ALM F30/7 by having Gln17 in place of Glu17, the latter occurring in the F30 group. Notably. currently commercially available alamethicins (Fluka, Sigma) represent microheterogeneous mixtures of the neutral ALM F50 peptides with trace amounts of acidic ALM F30 peptides.

To date, 18 genera of imperfect and ascomycetous fungi have been recognized to produce ca. 700 individual sequences of peptaibiotics. These are linear polypeptide antibiotics which i) have a molecular weight between 500 and 2,200 Dalton, thus containing 5-21 residues; ii) show a high content of alpha-aminoisobutyric acid; iii) are characterized by the presence of other nonproteinogenic amino acids and/or lipoamino acids; iv) possess an acylated N-terminus, and v) have a C-terminal residue that, in most of them, consists of a free or acetylated amide-bonded 1,2-amino alcohol, but might also be an amine, amide, free amino acid, 2,5-dioxopiperazine, or sugar alcohol. From April 2003 until present, ca. 300 new individual sequences of peptaibiotics have been published in the literature, but most of them have not yet been included in databases. To summarize these new sequences and novel constituents, as well as to introduce fungal species hitherto unknown as producers of peptaibiotics, the relevant literature is reviewed. Furthermore, ecophysiological and taxonomic aspects of the producing fungi are discussed.