title: Performance evaluation on a wide set of matrix-assisted laser desorption ionization matrices for the detection of oligosaccharides in a high-throughput mass spectrometric screening of carbohydrate depolymerizing enzymes.
abstract: Compared to other analytical methods, matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) presents several unique advantages for the structural characterization of degradation products of carbohydrates. Our final goal is to implement this technique as a high-throughput platform, with the aim of exploring natural bio-diversity to discover new carbohydrate depolymerizing enzymes. In this approach, a variety of carbohydrates will be used as enzymes substrates and MALDI-MS will be employed to monitor the oligosaccharides produced. One drawback of MALDI, however, is that the choice of the matrix is largely dependent on the chemical properties of the analyte. In this context, our objective in the present work was to find the smallest set of MALDI matrices able to detect chemically heterogeneous oligosaccharides. This was done through the performance evaluation of more than 40 MALDI matrices preparations. Homogeneity of analyte-matrix deposits was considered as a critical feature, especially since the final objective is to fully automate the analyses. Evaluation of the matrices was done by means of a rigorous statistical approach. Amongst all tested compounds, our work proposes the use of the DHB/DMA ionic matrix as the most generic matrix, for rapid detection of a variety of polysaccharides including neutral, anionic, methylated, sulfated, and acetylated compounds. The selected matrices were then used to screen crude bacterial incubation media for the detection of enzymatic degradation products.
authors.affiliation: INRA UR1268 Biopolymères Interactions Assemblages, Plate-Forme BIBS, F-44316 Nantes, France. David.Ropartz@nantes.inra.fr
authors.forename: David
authors.initials: D
authors.lastname: Ropartz
authors.forename: Pierre-Edouard
authors.initials: PE
authors.lastname: Bodet
authors.forename: Cédric
authors.initials: C
authors.lastname: Przybylski
authors.forename: Florence
authors.initials: F
authors.lastname: Gonnet
authors.forename: Régis
authors.initials: R
authors.lastname: Daniel
authors.forename: Maude
authors.initials: M
authors.lastname: Fer
authors.forename: William
authors.initials: W
authors.lastname: Helbert
authors.forename: Dominique
authors.initials: D
authors.lastname: Bertrand
authors.forename: Hélène
authors.initials: H
authors.lastname: Rogniaux
mesh_terms: D002457:Cell Extracts / Q000737:chemistry; D005690:Galactose / Q000031:analogs & derivatives; D006026:Glycoside Hydrolases / Q000737:chemistry* / Q000378:metabolism; D006247:Harmine / Q000737:chemistry; D057166:High-Throughput Screening Assays / Q000379:methods*; D062385:Hydroxybenzoates / Q000737:chemistry; D008190:Lyases / Q000737:chemistry / Q000378:metabolism; D008351:Mannans / Q000737:chemistry; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D031161:Pseudoalteromonas / Q000737:chemistry* / Q000378:metabolism; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000295:instrumentation*
chemical_list: D002457:Cell Extracts; D062385:Hydroxybenzoates; D008351:Mannans; D009844:Oligosaccharides; C012990:galactomannan; D006247:Harmine; D006026:Glycoside Hydrolases; D008190:Lyases; D005690:Galactose