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Author:
Date:
2014-05-05
[Abstract] Amino acid racemases are enzymes that invert the α-carbon stereochemistry of amino acids (AAs), interconverting amino acids between their L- and D-enantiomers in a reversible reaction. In bacteria, they are known to have catabolic physiological functions but are also involved in the synthesis of many D-AAs, including D-glutamate and D-alanine, which are necessary components of the peptidoglycan layer of the bacterial cell wall. As such, amino acid racemases represent significant targets for the development of bactericidal compounds. Amino acid racemases are also regarded by the ...
[摘要] 氨基酸消旋酶是在可逆反应中反转氨基酸（AA）的α-碳立体化学，在它们的L-和D-对映异构体之间相互转化氨基酸的酶。在细菌中，已知它们具有分解代谢的生理功能，但也参与许多D-AAs的合成，包括D-谷氨酸和D-丙氨酸，它们是细菌细胞壁的肽聚糖层的必需组分。因此，氨基酸消旋酶代表了杀菌化合物开发的重要目标。氨基酸消旋酶也被生物技术工业视为用于生产经济相关D-AAs的重要催化剂。在这里，我们提供使用高效液相色谱法（HPLC）和1-氟-2,4-二硝基苯基-5-L-丙氨酰胺（FDAA，也是Marfey的试剂）的新型氨基酸消旋酶的表征的详细协议。本文所述的方案被设计为获得准确的动力学参数（k cat cat，K M max值）。优化酶浓度和反应时间，以使反向反应最小化，这可在测定消旋酶反应时混淆结果。
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