Screening of sugar converting enzymes using quantitative MALDI-ToF mass spectrometry

被引：32
作者：
Bungert, D [1 ]
Bastian, S [1 ]
Heckmann-Pohl, DM [1 ]
Giffhorn, F [1 ]
Heinzle, E [1 ]
Tholey, A [1 ]
机构：
[1] Univ Saarland, D-66123 Saarbrucken, Germany
来源：
BIOTECHNOLOGY LETTERS | 2004年 / 26卷 / 13期
关键词：
ionic liquid matrices; MALDI-ToF MS; pyranose oxidase; quantitation; screening;
D O I：
10.1023/B:BILE.0000032965.18721.62
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
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.
引用
收藏
页码：1025 / 1030
页数：6
相关论文
共 13 条
  • [1] Quantitative matrix-assisted laser desorption/ionization mass spectrometry for the determination of enzyme activities
    Bungert, D
    Heinzle, E
    Tholey, A
    [J]. ANALYTICAL BIOCHEMISTRY, 2004, 326 (02) : 167 - 175
  • [2] A strategy for the determination of enzyme kinetics using electrospray ionization with an ion trap mass spectrometer
    Ge, X
    Sirich, TL
    Beyer, MK
    Desaire, H
    Leary, JA
    [J]. ANALYTICAL CHEMISTRY, 2001, 73 (21) : 5078 - 5082
  • [3] Fungal pyranose oxidases: occurrence, properties and biotechnical applications in carbohydrate chemistry
    Giffhorn, F
    [J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2000, 54 (06) : 727 - 740
  • [4] Kang MJ, 2000, RAPID COMMUN MASS SP, V14, P1972, DOI 10.1002/1097-0231(20001115)14:21<1972::AID-RCM119>3.0.CO
  • [5] 2-5
  • [6] Application of automated matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for the measurement of enzyme activities
    Kang, MJ
    Tholey, A
    Heinzle, E
    [J]. RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 2001, 15 (15) : 1327 - 1333
  • [7] Detection of protease activities with the mass-spectrometry-assisted enzyme-screening (MES) system
    Schlüter, H
    Jankowski, J
    Rykl, J
    Thiemann, J
    Belgardt, S
    Zidek, W
    Wittmann, B
    Pohl, T
    [J]. ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2003, 377 (7-8) : 1102 - 1107
  • [8] Second-generation MS-based high-throughput screening system for enantioselective catalysts and biocatalysts
    Schrader, W
    Eipper, A
    Pugh, DJ
    Reetz, MT
    [J]. CANADIAN JOURNAL OF CHEMISTRY, 2002, 80 (06) : 626 - 632
  • [9] The production of fine chemicals by biotransformations
    Straathof, AJJ
    Panke, S
    Schmid, A
    [J]. CURRENT OPINION IN BIOTECHNOLOGY, 2002, 13 (06) : 548 - 556
  • [10] Derivatization of small biomolecules for optimized matrix-assisted laser desorption/ionization mass spectrometry
    Tholey, A
    Wittmann, C
    Kang, MJ
    Bungert, D
    Hollemeyer, K
    Heinzle, E
    [J]. JOURNAL OF MASS SPECTROMETRY, 2002, 37 (09): : 963 - 973
12