The biocatalytic oxidative deamination- reductive amination process for the production of L-phosphinothricin (L-PPT) from D,L-phosphinothricin (D,L-PPT) is a green and environmentally friendly approach with significant development potential. In this study, the adopted technological route involves the oxidative deamination of the D-PPT component in D,L-PPT to 2-oxo-4-(hydroxymethylphosphinyl)butanoic acid (PPO) by utilizing D-amino acid oxidase (DAAO), followed by the catalytic reductive amination of PPO to L-PPT using phosphinothricin dehydrogenase (PpDH). In order to enhance the catalytic rate and reduce the inhibitory effect of the intermediate product PPO on the catalytic process, optimization of both oxidative deamination and reductive amination phases was conducted through separate strategies of constant-pressure oxygen supply and substrate feeding, respectively. A “two-step one-pot” method was employed to link the optimized reactions, significantly shortening the production time. Furthermore, the “two-step one-pot” catalytic strategy was optimized, ultimately achieving a high 97.7% yield of L-PPT with ee of L-PPT>99.9% after 5.5 h of reaction under 800 mM D,L-PPT catalytic conditions, D-PPT and PPO could not bedetected in the reaction system, further enchanced the catalytic efficiency, production efficiency and product quality. The aforementioned results demonstrate that this proposed approach has great potential for industrial-scale production of L-PPT as well as significant competitive advantages in terms of economic feasibility.

生物催化氧化脱氨-还原氨化工艺由D,L-草胺膦( D, L -PPT)生产L-草胺膦(L-PPT)是一种绿色环保的方法，具有巨大的发展潜力。本研究采用的技术路线是利用D-氨基酸氧化酶(DAAO)将D , L -PPT中的D -PPT组分氧化脱氨基为2-氧代-4-(羟甲基次膦酰)丁酸(PPO)，然后使用膦丝菌素脱氢酶 (PpDH) 将 PPO 催化还原胺化为L -PPT。为了提高催化速率并减少中间产物PPO对催化过程的抑制作用，分别通过恒压供氧和底物进料策略对氧化脱氨和还原胺化阶段进行优化。采用“两步一锅”方法链接优化反应，显着缩短生产时间。此外，优化了“两步一锅”催化策略，在800 mM D , L - 下反应5.5 h后，最终实现了L -PPT的高产率97.7%， ee为L -PPT>99.9%。 PPT催化条件下，反应体系中检测不到D-PPT和PPO，进一步提高了催化效率、生产效率和产品质量。 上述结果表明，该方法具有工业规模生产L -PPT的巨大潜力，并且在经济可行性方面具有显着的竞争优势。