Deracemization of D,L-phosphinothricin (D,L-PPT) is one of the most promising routes for preparation of optically pure L-PPT. In this work, an efficient multi-enzyme redox cascade was developed for deracemization of D,L-PPT, which includes oxidative reaction and reductive reaction. The oxidative reaction catalyzing oxidative deamination of D-PPT to 2-oxo-4-[(hydroxy)(-methyl)phosphinyl]butyric acid (PPO) was performed by a D-amino acid oxidase and a catalase for removing H₂O₂. The reductive reaction catalyzing amination of PPO to L-PPT is achieved by a glufosinate dehydrogenase and a glucose dehydrogenase for cofactor regeneration. To avoid the inhibitory effect of glucose on the oxidative reaction, a “two stages in one-pot” strategy was developed to combine these two reactions in deracemization process. By using this strategy, the L-PPT was obtained with a high yield (89%) and > 99% enantiomeric excess at substrate loading of 300 mM in absence of addition of extra NADP⁺. These encouraging results demonstrated that the developed enzyme cascade deracemization process exhibits great potential and economical competitiveness for manufacture of L-PPT from D,L-PPT.

D,L-膦丝菌素 (D,L-PPT) 的脱消旋是制备光学纯 L-PPT 的最有前途的途径之一。在这项工作中，开发了一种有效的多酶氧化还原级联反应，用于 D,L-PPT 的去消旋，包括氧化反应和还原反应。通过 D-氨基酸氧化酶和过氧化氢酶去除 H ₂ O ₂. 催化 PPO 胺化为 L-PPT 的还原反应是通过草铵膦脱氢酶和用于辅助因子再生的葡萄糖脱氢酶实现的。为了避免葡萄糖对氧化反应的抑制作用，开发了“一锅两步”策略，将这两个反应结合在去消旋过程中。通过使用该策略，在不添加额外 NADP ^{+ 的}情况下，在 300 mM 的底物负载下，以高产率 (89%) 和 > 99% 的对映体过量获得 L-PPT 。这些令人鼓舞的结果表明，所开发的酶级联去消旋工艺在从 D,L-PPT 制造 L-PPT 方面表现出巨大的潜力和经济竞争力。