title: Development and validation of LC-MS methods for peptaibol quantification in fungal extracts according to their lengths.
abstract: Some terrestrial Trichoderma sp. strains are already used as biological control agents (BCAs). They all produce peptaibols, small antimicrobial peptides which are supposed to play a role in the anti-phytopathogenic activity of Trichoderma sp. Trichoderma strains producing high amounts of peptaibols could represent new potential BCAs. In this context, marine-derived Trichoderma strains from the marine fungal strain collection of the "Mer, Molécules, Santé" (MMS) laboratory were investigated for their peptaibol production. Previously, the quantification of peptaibols was performed using alamethicin, as standard (20-amino acid residues peptaibol). In this study, the development and validation of quantification LC/ESI-TI-MS methods using different standards of peptaibols (11-, 14- and 20-amino acid residues) was performed in order to quantify all of them, in a single analysis, in Trichoderma crude extracts according to their chain length. The developed and validated methods were used to study the peptaibol production kinetic of a marine-derived Trichoderma strain, i.e., Trichoderma longibrachiatum (MMS 151). The results showed the optimal culture time at the 9th day with concentrations reaching 1.4±0.2% and 2.3±0.4% of the fungal biomass respectively for 11- and 20-residue peptaibols. Then, the different peptaibol subgroups produced by 13 Trichoderma strains were quantified. According to their 18-, 19- and 20-residue peptaibol production, three strains referenced as MMS 1541, MMS 639 and MMS 151 seemed to be good candidates as potential new biological control agents with respective production of 0.4, 0.4 and 2.1%.
authors.affiliation: University of Nantes, MMS, EA 2160, F-44000 Nantes, France.
authors.forename: Anne-Isaline
authors.initials: AI
authors.lastname: Van Bohemen
authors.affiliation: University of Nantes, MMS, EA 2160, F-44000 Nantes, France. Electronic address: aurore.zalouk-vergnoux@univ-nantes.fr.
authors.forename: Aurore
authors.initials: A
authors.lastname: Zalouk-Vergnoux
authors.affiliation: University of Nantes, MMS, EA 2160, F-44000 Nantes, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Poirier
authors.affiliation: University of Nantes, MMS, EA 2160, F-44000 Nantes, France.
authors.forename: Nam Ngoc
authors.initials: NN
authors.lastname: Phuong
authors.affiliation: UPVD, University of Perpignan, CRIOBE-USR CNRS 3278, F-66860 Perpignan, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Inguimbert
authors.affiliation: UPVD, University of Perpignan, CRIOBE-USR CNRS 3278, F-66860 Perpignan, France.
authors.forename: Khoubaib
authors.initials: K
authors.lastname: Ben Haj Salah
authors.affiliation: University of Nantes, MMS, EA 2160, F-44000 Nantes, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Ruiz
authors.affiliation: University of Nantes, MMS, EA 2160, F-44000 Nantes, France.
authors.forename: Yves François
authors.initials: YF
authors.lastname: Pouchus
mesh_terms: D000595:Amino Acid Sequence; D000890:Anti-Infective Agents / Q000032:analysis* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D057230:Limit of Detection; D008969:Molecular Sequence Data; D054848:Peptaibols / Q000032:analysis* / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D014242:Trichoderma / Q000737:chemistry* / Q000378:metabolism
chemical_list: D000890:Anti-Infective Agents; D054848:Peptaibols
keywords: Biocontrol; LC/ESI-IT-MS; Peptaibols; Quantification; Trichoderma sp; Validation