Quantitative matrix-assisted laser desorption/ionization mass spectrometry for the determination of enzyme activities

被引：43

作者：

Bungert, D ^{[1
]}

Heinzle, E ^{[1
]}

Tholey, A ^{[1
]}

机构：

[1] Univ Saarland, D-66123 Saarbrucken, Germany

来源：

ANALYTICAL BIOCHEMISTRY | 2004年 / 326卷 / 02期

关键词：

enzyme activity; quantification; low-molecular-weight compounds; screening;

D O I：

10.1016/j.ab.2003.11.013

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Quantitative matrix-assisted laser desorption/ionization (MALDI) time-of-flight (ToF) mass spectrometry (MS) was applied for the determination of concentrations of low-molecular-weight (<400 Da) substrates and products of enzyme-catalyzed reactions. Isotope-labeled and fluorinated internal standards were used for the quantification. Automated quantitative MALDI-ToF MS analysis of quenched samples allowed the direct and simultaneous observation of time-dependent decrease of substrate concentration and increase of product concentration without any need for prepurification or desalting steps. The results showed good agreement with established but more elaborate analytical methods. MALDI-ToF MS thus is an interesting alternative tool for the determination of enzyme activities. Due to automated and miniaturized measurement it is especially suitable for the screening of biocatalysts. (C) 2003 Elsevier Inc. All rights reserved.

引用

页码：167 / 175

页数：9

共 43 条

[1]

[Anonymous], 1972, The Enzymes

[2] Monitoring enzyme catalysis with mass spectrometry [J].