Figure 4. Regio- and enantioselective multiple oxy- and amino-functionalizations of styrene 1a with E. coli strains containing multiple enzyme modules.

(a) Product concentration of biotransformation of 100 mM 1a to (S)-5a with twelve E. coli strains (10 g cdw l⁻¹), each containing both enzyme module 1 and 2, respectively. (b) Time course of biotransformation of 120 mM 1a to (S)-5a with E. coli (A-M1_R-M2) cells (15 g cdw l⁻¹) in a two-liquid-phase system (KP buffer containing 0.25% glucose and n-hexadecane; 1:1) at 30 °C. (c) Product concentration of biotransformation of 50 mM 1a to (S)-6a with twelve E. coli strains (10 g cdw l⁻¹), each containing both enzyme module 1 and 3, respectively. (d) Time course of biotransformation of 60 mM 1a to (S)-6a with E. coli (A-M1_E-M3) cells (15 g cdw l⁻¹) in a two-liquid-phase system (NaP buffer containing 1% glucose and 200 mM NH₃/NH₄Cl and n-hexadecane; 1:1) at 25 °C. (e) Product concentration of biotransformation of 50 mM 1a to (S)-8a with eight E. coli strains (10 g cdw l⁻¹), each containing enzyme module 1, 2 and 4, respectively. (f) Time course of biotransformation of 60 mM 1a to (S)-8a with E. coli (A-M1_R-M2_C-M4) cells (15 g cdw l⁻¹) in a two-liquid-phase system (KP buffer containing 0.5% glucose and 100 mM NH₃/NH₄Cl and n-hexadecane; 1:1) at 30 °C (arrow: adding additional 0.5% glucose and 100 mM NH₃/NH₄Cl at 20 h). All biotransformations were performed in triplicate, and error bars show±s.d.