Figure 3.

(A) Substrate scope of Sav HppD Az1 and Sav HppD Az2. Experiments were performed at analytical scale using suspensions of E. coli expressing Sav HppD variants in KPi buffer (pH 7.4) at room temperature under anaerobic conditions for 24 hours (detailed conditions see Table S3). The absolute configuration of enzymatically synthesized azidation product 1 was determined to be S via X-ray crystallography. The absolute configurations of all other azidation products were inferred by analogy. (B) Preparative scale synthesis and absolute configuration determination. (C) One-pot chemoenzymatic synthesis by in situ derivatization of enzymatic azidation products via CuAAC. Detailed conditions see section IX of the SI.