TAILIEUCHUNG - Báo cáo khoa học: Direct identification of hydrophobins and their processing in Trichoderma using intact-cell MALDI-TOF MS

Intact-cell MS (ICMS) was applied for the direct detection of hydropho-bins in various species and strains ofHypocrea⁄ both myce-lia and spores, dominating peaks were identified as hydrophobins by detecting mass shifts of 8 Da of reduced and unreduced forms, the analysis of knockout mutants, and comparison with protein databases. | ỊFEBS Journal Direct identification of hydrophobins and their processing in Trichoderma using intact-cell MALDI-TOF MS Torsten Neuhof1 Ralf Dieckmann1 Irina S. Druzhinina2 Christian P. Kubicek2 Tiina Nakari-Setala3 Merja Penttila3 and Hans von Dohren1 1 TU Berlin Institut fur Chemie FG Biochemie und Molekulare Biologie Berlin Germany 2 FB Gentechnik und Angewandte Biochemie Institut fur Verfahrenstechnik Umwelttechnik und Technische Biowissenschaften TU Wien Vienna Austria 3 VTT TechnicalResearch Centre of Finland Espoo Finland Keywords fungalbiomarker hydrophobin intact-cell MS MALDI-TOF MS Trichoderma Correspondence H. von Dohren TU Berlin Institut fur Chemie FG Biochemie und Molekulare Biologie Franklinstr. 29 10587 Berlin Germany Fax 49 30 314 24783 Tel 49 30 314 22697 E-mail Doehren@ Present address AnagnosTec Gesellschaft fur Analytische Biochemie und Diagnostik mbH Potsdam-Golm Germany Intact-cell MS ICMS was applied for the direct detection of hydrophobins in various species and strains of Hypocrea Trichoderma. In both myce-lia and spores dominating peaks were identified as hydrophobins by detecting mass shifts of 8 Da of reduced and unreduced forms the analysis of knockout mutants and comparison with protein databases. Strain-specific processing was observed in the case of Hypocrea jecorina anamorph Trichoderma reesei . An analysis of 32 strains comprising 29 different species of Trichoderma and Hypocrea showed hydrophobin patterns that were specific at both at the species and isolate subspecies levels. The method therefore permits rapid and direct detection of hydrophobin class II compositions and may also provide a means to identify Trichoderma and other fungal species and strains from microgram amounts of biomass without prior cultivation. Received 19 September 2006 revised 27 November 2006 accepted 6 December 2006 doi Hydrophobins are small proteins thought to be ubiquitous in filamentous fungi. They .

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