非表示:
キーワード:
multiplexed ESI-MS; high-throughput screening; enantioselectivity; combinatorial catalysis; directed evolution
要旨:
A high-throughput method is described, where the enantioselectivity of approximately 10 000 catalysts or biocatalysts can be determined per day. The method is based on electrospray mass spectrometric techniques using an eight- channel multiplexed (MUX) sprayer system connected to a time- of-flight mass spectrometer. The inlet of the ion source is controlled by a stepping rotor that is continuously moving from one sprayer to the next with a recording time of 100 ms for each channel and a delay time of 50 ms, thus allowing a spectrum to be obtained from each channel every 1.2 s. One cycle, where eight samples are being sprayed in parallel, requires around 70 s, which allows a 96-well microtiter plate to be screened in 14 min. Integration of two pseudo-enantiomers (S)-glycidyl phenyl ether and (R)-D-5-glycidyl phenyl ether is necessary to quantify the enantiomeric excess (ee-value), where one enantiomer is isotopically labeled to allow easy identification of the mass spectrometric signals. Errors of similar to2% for the ee-values indicate that in addition to the significant improvement in sample throughput this is also a precise method for high-throughput screening. This second- generation assay is useful for combinatorial enantioselective transition-metal catalysis and in the directed evolution of enantioselective enzymes.
