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The reaction mixtures (100 L) contained 1.0 M Cp1B, 10 mM ATP, 12 mM MgCl 2 , 300 M NADH, 500 M phosphoenolpyruvic acid (PEP), 41 units/mL pyruvate kinase (PK, Sigma), 59 units/mL lactate dehydrogenase (LDH, Sigma), 10 mM KCl and 100 mM Tris-HCl (pH 8.0) with various concentration (0.04 mM to 1 mM) of (2 S ,3 S )- t -ES and 5 mM l-Phe