NMR as a readout to monitor and restore the integrity of complex chemoenzymatic reactions.

NMR as a readout to monitor and restore the integrity of complex chemoenzymatic reactions.
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NMR作为监测和恢复复杂化学酶反应的完整性的读数。

DOI:
10.1016/j.jmr.2022.107265
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发表时间:
2022-09
影响因子:
2.2
通讯作者:
Frueh, Dominique P.
Frueh, Dominique P.
中科院分区:
化学3区
文献类型:
--
作者:
Marincin, Kenneth A.;Hwang, Yousang;Kengmana, Everett S.;Meyers, David J.;Frueh, Dominique P.

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核磁共振的非侵入性为在原子水平上监测生物化学反应提供了一种手段。我们利用这一优势来监测复杂的化学酶反应,该反应顺序修饰试剂并将产物装载到非核糖体肽合成酶载体蛋白上。我们提出了一个包括脉冲序列的方案,该方案允许评估试剂的完整性和反应中每个步骤的完成情况,从而减轻了耗时和昂贵的方法来调试和重复低效的反应。这项研究强调了核磁共振作为建立可靠和可重复的生化研究实验条件的工具的重要性。
The non-invasive nature of NMR offers a means to monitor biochemical reactions in situ at the atomic level. We harness this advantage to monitor a complex chemoenzymatic reaction that sequentially modifies reagents and loads the product on a nonribosomal peptide synthetase carrier protein. We present a protocol including a pulse sequence that permits to assess both the integrity of reagents and the completion of each step in the reaction, thus alleviating otherwise time-consuming and costly approaches to debug and repeat inefficient reactions. This study highlights the importance of NMR as a tool to establish reliable and reproducible experimental conditions in biochemical studies.
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