The synthesis of substituted d-phenylalanines in high yield and excellent optical purity, starting from inexpensive cinnamic acids, has been achieved with a novel one-pot approach by coupling phenylalanine ammonia lyase (PAL) amination with a chemoenzymatic deracemization (based on stereoselective oxidation and nonselective reduction). A simple high-throughput solid-phase screening method has also been developed to identify PALs with higher rates of formation of non-natural d-phenylalanines. The best variants were exploited in the chemoenzymatic cascade, thus increasing the yield and ee value of the d-configured product. Furthermore, the system was extended to the preparation of those l-phenylalanines which are obtained with a low ee value using PAL amination.
Phenylalanin‐Ammoniak‐Lyase (PAL) wurde mit l‐Aminosäure‐Desaminase (LAAD) oder d‐Aminosäure‐Oxidase (DAAO) zu einer Kaskade gekoppelt, um preiswert verfügbare Zimtsäuren chemoenzymatisch in nichtnatürliche d‐ und l‐Phenylalanine umzuwandeln. Ein neuartiges Festphasen‐Screening führte zu PAL‐Varianten mit erhöhter d‐Aktivität, die verbesserte Ausbeuten und ee‐Werte ergaben.WILEY-VCH.
Keywords: Aminosäuren; Biokatalyse; Enzyme; Lyasen; Synthesemethoden.