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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Optimization of sample preparation for intact cell mass spectrometry (matrix-assisted laser desorption/ionization linear time-of-flight mass spectrometry) of endophytic Xylaria

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Author(s):
Amaral, Luciana da Silva [1, 2] ; Rodrigues-Filho, Edson [1] ; Kubicek, Christian P. [3, 4] ; Herwig, Christoph [3] ; Marchetti-Deschmann, Martina [2] ; Allmaier, Guenter [2]
Total Authors: 6
Affiliation:
[1] Univ Fed Sao Carlos, Dept Quim, CP 676, BR-13565905 Sao Carlos, SP - Brazil
[2] Vienna Univ Technol, TU Wien, Inst Chem Technol & Analyt, Getreidemarkt 9-164, A-1060 Vienna - Austria
[3] Vienna Univ Technol, TU Wien, Inst Chem Environm & Biosci Engn, Getreidemarkt 9-166, A-1060 Vienna - Austria
[4] Steinschotelgasse 7, A-1100 Vienna - Austria
Total Affiliations: 4
Document type: Journal article
Source: RAPID COMMUNICATIONS IN MASS SPECTROMETRY; v. 32, n. 10, p. 815-823, MAY 30 2018.
Web of Science Citations: 0
Abstract

RationaleAlthough the fruiting-body of the fungi of the genus Xylaria shows a great variety of morphological characteristics, their mycelial forms are always very similar, imposing difficulties for their identification. Intact cell mass spectrometry (ICMS) using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) can be a fast and reliable strategy to support the differentiation/identification of Xylaria species in those cases where fruit-bodies are not available. MethodsMany experimental parameters such as sample preparation and culture media are crucial for filamentous fungi analysis by MALDI-TOFMS. For the purposes of this study, we used four matrices (CHCA, DHB, FA and SA) with five different concentrations (0.1, 0.3, 0.5, 1.0 and 2.5%) of TFA in the matrix, the influence of six different culture media (solid and liquid), and three mycelium peptide/protein extraction protocols (acid, basic and thymol-supported solution) to optimize the sample preparation of the endophytic fungus X. arbuscula. ResultsIt was observed that sinapinic acid (30mg/mL) dissolved in acetonitrile/0.1% TFA and PDA were the best matrix solution and culture medium, respectively, for the ICMS of X. arbuscula. The formic acid and ammonium bicarbonate (AB) protocols provided similar mass spectra; however, a higher number of peaks were observed using AB extraction. Mass spectra obtained from different thymol-containing solutions (EtOH/aqueous 0.1% TFA and ACN/aqueous 0.1% TFA) show increasing peak abundances at m/z 3000-6500. ConclusionsX. arbuscula could be analyzed by ICMS. However, an extraction step was required to provide suitable MALDI mass spectra. Formic acid-, AB- and thymol-containing solutions were demonstrated to be good cocktails for the extraction of peptide/protein biomarkers from these fungi. (AU)

FAPESP's process: 13/18603-3 - Metabolic and biosynthetic studies for the chemical and taxonomic contribution of Xylariaceae family microorganisms
Grantee:Luciana da Silva Amaral
Support type: Scholarships in Brazil - Doctorate