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With phenylalanine ammonia-lyase from Pseudozyma antarctica yeast; In aq. buffer; at 30℃; for 168h;pH 8.5;Resolution of racemate;
General procedure: The ammonia elimination reaction mixtures containing 5mM of theracemic phenylalanines (rac-1a-s) in 100mM TRIS buffer (pH 8.5) andpurified PzaPAL or PcPAL (50 Xg, either) in 1 mL reaction volume wereincubated at 30 C. Samples (50 XL) taken at different time points (17,40, 64, and 168 h) from the reaction mixtures were analyzed by HPLCfor conversion and ee values, using previously developed methods [44].The ammonia addition and elimination reactions were followed byHPLC measurements. Conversions were determined on Agilent 1200HPLC instrument using Phenomenex Gemini NX-C-18 column and amixture of NH4OH buffer (0.1 M, pH 8.5) and MeOH at a flow rate of1 mL min-1. Conversions were calculated from peak area integrationswith use of appropriate response factor (Table S2). Enantiomer separationswere carried out on Agilent 1100 HPLC instrument usingCrownpak CR-I (+) column and a mixture of aqueous HClO4, pH 1.5and acetonitrile as eluent at a flow rate of 0.4 mL min-1. (Table S3).