Screening of sugar converting enzymes using quantitative MALDI-ToF mass spectrometry.
Quantitative matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-ToF MS) was applied for the screening of ten pyranose oxidase variants. Quantitative MALDI-ToF MS using isotopic labeled internal standards and ionic liquid matrices was performed using aliquots of enzyme reaction mixtures without prepurification steps. The results obtained were in good agreement with HPLC measurements. Analysis time was approx. 3.5 min for a five-fold determination. Thus, quantitative MALDI-ToF MS can be used as a tool for screening of sugar converting enzymes.
Bungert
Ditte
D
Technische Biochemie, Universität des Saarlandes, 66123 Saarbrücken, Germany.
Bastian
Sabine
S
Heckmann-Pohl
Dorothée M
DM
Giffhorn
Friedrich
F
Heinzle
Elmar
E
Tholey
Andreas
A
D002237:Carbohydrate Dehydrogenases / Q000737:chemistry / Q000378:metabolism*; D050260:Carbohydrate Metabolism; D002851:Chromatography, High Pressure Liquid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D002237:Carbohydrate Dehydrogenases; C022489:pyranose oxidase