Derivatization using dimethylamine for tandem mass spectrometric structure analysis of enzymatically and acidically depolymerized methyl cellulose.
Structure analysis of partially depolymerized methyl cellulose was performed by nanoelectrospray ionization tandem mass spectrometry (nano-ESI-MS/MS) and by matrix-assisted laser desorption/ionization tandem mass spectrometry (MALDI-MS/MS). Dimethylamine (DMA) was used for the first time as a reducing end derivatization reagent for oligosaccharides. This is an attractive reagent since it could be easily removed from the reaction mixture. Most important it also introduces a basic functional group that increased the sensitivity in both MALDI and nano-ESI. Depolymerization was made in two ways: one by the cellulose selective endoglucanase 5A from Bacillus agaradhaerens (Ba Cel5A) and the other by trifluoroacetic acid. The DMA derivatives formed both protonated and sodiated molecules in nano-ESI and MALDI. Tandem MS of protonated molecules yielded predominantly Y fragments from which the distribution of the substituents in the oligomers could be measured. Fragments obtained in tandem MS of sodiated molecules provided information regarding the positions of the substituents within the anhydroglucose units (AGUs). It was found that Ba Cel5A could cleave glucosidic bonds also if the AGU on the reducing side of the bond was fully methylated. The combination of DMA derivatization and tandem MS was demonstrated as a tool for the characterization of endoglucanase selectivity.
Momcilovic
Dane
D
Department of Technical Analytical Chemistry, Lund University, P.O. Box 124, S-221 00 Lund, Sweden.
Schagerlöf
Herje
H
Röme
Daniel
D
Jörntén-Karlsson
Magnus
M
Karlsson
Karl-Erik
KE
Wittgren
Bengt
B
Tjerneld
Folke
F
Wahlund
Karl-Gustav
KG
Brinkmalm
Gunnar
G
D002480:Cellulase / Q000737:chemistry / Q000378:metabolism; D004123:Dimethylamines / Q000737:chemistry*; D008747:Methylcellulose / Q000737:chemistry* / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014269:Trifluoroacetic Acid / Q000737:chemistry
D004123:Dimethylamines; D008747:Methylcellulose; C034516:dimethylamine; D014269:Trifluoroacetic Acid; C407024:endoglucanase Cel5A; D002480:Cellulase