LCMS/MS-Based Mining of the Peptaibiome: A Discovery of New Peptaibols

In this study, we employed liquid chromatography–tandem
mass spectrometry (LC–MS/MS) to perform chemo-profiling on
four fungal species: three Trichoderma speciestwo strains
of Trichoderma asperellum (S3–3 and S3–1)
and one Trichoderma longibrachiatum (FS-46)and
one species from the Tolypocladium genus, Tolypocladium inflatum (MTCC-3538). Our analysis identified
78 peptaibols featuring α-aminoisobutyric acid (Aib). Among
these, we discovered both cyclic and linear Peptaibols and lipopeptaibols
with Aib, ranging in putative molecular weights from 1033.4 to 1953.6
g/mol, corresponding to peptide lengths of 10–20 amino acids.
In total, 78 peptaibols, including lipopeptaibols, were identified
through LC-MS/MS. Of these, 43 were recognized as new peptaibols based
on mass fragmentation patterns and sequence data from the MS/MS analysis.
Our findings suggest that LC-MS/MS-based large-scale studies of the
peptaibiome will enable the discovery of novel peptaibols and provide
a valuable reference for understanding fragmentation patterns of peptaibols,
cyclic peptaibols, and lipopeptaibols. Moreover, LC-MS/MS allows rapid
identification of peptaibiotics at low concentrations directly in
extracts, offering a potential quality control method for selecting
beneficial Trichoderma strains for crop protection, while their fragmentation
patterns and plausible sequences can guide synthetic design.