Chloramphenicol → (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal

Participants

Reaction Steps

For inoculum preparation, the E. coli MG1655:: pCECNde/4978 DAT/SEQ ID NO:172 were grown overnight at 37° C. with shaking at 250 rpm in Luria-Bertani (“1.13”) medium with 25 μg/mL chloramphenicol. For the experimental treatments, Trp-1+glucose medium, a minimal medium that has been used for increased production of tryptophan in E. coli cells (Zeman et al. Folia Microbiol. 35:200-4, 1990), was prepared as follows. To 800 mL nanopure water the following reagents were added: 2 g (NH4)2SO4. 13.6 g KH2PO4. The pH was adjusted to 7.0, the volume was increased to 948 mL, and the medium was autoclaved. Following sterilization, 0.2 g MgSO4.7H2O, 0.01 g CaCl2′2H2O, and 0.5 mg FeSO4.7H2O were added to the medium in a 1.8 mL volume followed by addition of 0.2 mL of Neidhardt's micronutrient solution (Neidhardt F. C., Bloch P. L., and Smith D. F., 1974. Culture medium for Enterobacteria. J. Bacteriol. 119: 736-746). Neidhardt's medium includes (per liter): 0.18 g (NH4)6(MO7)24.4H2O, 1.24 g H3BO3, 0.36 g CoCl2.6H2O, 0.12 g CuSO4 (anhydrous), 0.8 g MnCl2.4H2O, and 0.14 g ZnSO4.7H2O. A 50% glucose solution was prepared separately and sterile-filtered. Forty mL of glucose solution and 10 mL of 1 M 3-morpholinopropanesulfonic acid (“MOPS”) buffer were added to the base medium (950 mL) for a 1 L final volume.

Provenance & Evidence