Fig. 4: Interfacing the Lossen rearrangement with native and engineered metabolic pathways.
From: A biocompatible Lossen rearrangement in Escherichia coli
a, Interfacing the Lossen rearrangement with native and engineered biosynthetic pathways in E. coli. b, Biotransformation of DMM (5) to dimethylsuccinate (6) and β-ketoacrylate 7 to γ-ketoester 8 using E. coli BW25113∆pabB and 1 or PET-1. c, A de novo biosynthetic pathway to 4-AP (9) and paracetamol 10 incorporating a non-enzymatic Lossen rearrangement, plasmid designs and whole-cell production experiments. d, Paracetamol synthesis by one-pot Lossen rearrangement and bacterial whole-cell synthesis. Strain E. coli_p350 expresses ABH60 and strain E. coli_p354 expresses PANATK211G. Ratios refer to E. coli_p354 and E. coli_p350 in biotransformations (1:1, OD600 26; 1:5, OD600 16; 1:100 and 1:200, OD600 12.5). aFinal cell density OD600 25. OD refers to the final optical density at 600 nm. bCells induced with 0.5% l-arabinose. Reaction conditions: (i) substrate (0.5 mM), aqueous potassium phosphate (200 mM, pH 8.0), 50 °C, 48 h; (ii) E. coli_p354 and E. coli_p350 (1:10; OD600 20). Biotransformations were analysed by 1H NMR relative to an internal standard of trimethoxybenzene or by HPLC relative to an internal standard of caffeine. All data are presented as mean values ± s.d. of three biological replicates.
