Enzymatic Reactions Observed with Zero- and Low-Field Nuclear Magnetic Resonance
Enzymatic Reactions Observed with Zero- and Low-Field Nuclear Magnetic Resonance
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DOI：
10.1021/acs.analchem.3c02087
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发表时间：
2023-12-04
影响因子：
7.4
通讯作者：
Blanchard，John W.
中科院分区：
文献类型：
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作者：
Eills，James;Picazo-Frutos，Roman;Blanchard，John W.
我们证明，酶催化反应可以观察到在零场和低场NMR实验结合最新进展仲氢为基础的超极化方法与国家的最先进的磁力。具体而言，我们研究了两个模型的生物过程：富马酸转化为苹果酸，这是在体内作为细胞坏死的标志物，和丙酮酸转化为乳酸，这是最广泛研究的代谢过程中超极化增强成像。除此之外，我们还构建了一个微流控零场NMR装置，用于在芯片实验室设备中对[1- 13 C]富马酸盐的微升规模样品进行实验。零至超低场（ZULF）NMR相对于高场NMR具有两个关键优势：信号可以穿过导电材料（例如，金属），并且来自样品不均匀性的谱线展宽可忽略不计。迄今为止，使用ZULF NMR进行工艺监测仅限于研究氢化反应。在这项工作中，我们展示了这种新兴的分析技术，更一般的反应监测和比较零与低场检测。
We demonstrate that enzyme-catalyzed reactions can be observed in zero- and low-field NMR experiments by combining recent advances in parahydrogen-based hyperpolarization methods with state-of-the-art magnetometry. Specifically, we investigated two model biological processes: the conversion of fumarate into malate, which is used in vivo as a marker of cell necrosis, and the conversion of pyruvate into lactate, which is the most widely studied metabolic process in hyperpolarization-enhanced imaging. In addition to this, we constructed a microfluidic zero-field NMR setup to perform experiments on microliter-scale samples of [1-13C]fumarate in a lab-on-a-chip device. Zero- to ultralow-field (ZULF) NMR has two key advantages over high-field NMR: the signals can pass through conductive materials (e.g., metals), and line broadening from sample heterogeneity is negligible. To date, the use of ZULF NMR for process monitoring has been limited to studying hydrogenation reactions. In this work, we demonstrate this emerging analytical technique for more general reaction monitoring and compare zero- vs low-field detection.