AbstractTrichoderma spp. are regularly found as a constituent of the mycoflora of many soils and are noted for their antagonistic activity against bacteria and other fungi. This latter property is the basis for the widespread interest in their use in the biological control of soil‐borne fungal plant pathogens. This antagonism is partly based on their ability to produce an impressive inventory of secondary metabolites. An important group of bioactive metabolites produced by Trichoderma spp. are the non‐ribosomal peptides (NRPs), especially the peptaibols. A virulent antagonistic strain, T. asperellum, which had been used in biological control strategies in Malaysia and previously examined for mycolytic enzyme production, has been studied for its potential for peptaibol production. The present research demonstrated the ability of T. asperellum to produce at least two metabolites which were identified as acid trichotoxin 1704E (Ac‐Aib‐Gly‐Aib‐Leu‐Aib‐Gln‐Aib‐Aib‐Aib‐Ala‐Ala‐Aib‐Pro‐Leu‐Aib‐Iva‐Glu‐Vol) and neutral trichotoxin 1717A (Ac‐Aib‐Gly‐Aib‐Leu‐Aib‐Gln‐Aib‐Aib‐Aib‐Ala‐Aib‐Aib‐Pro‐Leu‐Aib‐Iva‐Gln‐Vol). Addition of free Aib to the culture medium enhanced the production of trichotoxins. Biological activity of these substances was investigated against Bacillus stearothermophilus. The general characteristics of peptaibols, also found in the trichotoxins, include the presence of high proportions of the uncommon amino acid Aib, the protection of the N‐ and C‐termini by an acetyl group and reduction of the C‐terminus to 2‐amino alcohols, respectively, amphipathy and microheterogeneity.