Characterization of enzymatic pectin digests by matrix-assisted laser desorption/ionization mass spectrometry.
The use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for the characterization of partially methyl-esterified enzymatic pectin digests is described. The sensitivities of several matrices, positive and negative ion modes and desalting techniques for these acidic oligosaccharides were compared. The most favorable results were obtained with a thin-layer preparation of a mixture of 2,4,6-trihydroxyacetophenone and nitrocellulose in the negative ion mode. Results are presented demonstrating the sensitive characterization of separated and unseparated high-ester pectin digests obtained after complete digestion using Aspergillus niger pectin lyase and the analysis of digests after chemical modification. In the case of unseparated digests, the analysis of methylation patterns is demonstrated. Oligomers with a degree of polymerization up to 40 were detected after enrichment of large oligomers by propan-2-ol precipitation.
Odense University, Department of Molecular Biology, Denmark.
R
R
Körner
G
G
Limberg
J D
JD
Mikkelsen
P
P
Roepstorff
D000433:1-Propanol; D011232:Chemical Precipitation; D002852:Chromatography, Ion Exchange; D008745:Methylation; D011133:Polysaccharide-Lyases / Q000378:metabolism*; D013048:Specimen Handling / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D000433:1-Propanol; D011133:Polysaccharide-Lyases; C027349:pectin lyase