Ampicillin → L-Phenylalanine

Participants

Reaction Steps

L-phenylalanine was synthesized using cells of E. coli RR1 containing the plasmid pBPDH3. As an NADH regeneration system, enzymes for glycolysis in E. coli were used. The above-mentioned E. coli was inoculated in an LB medium containing 50 μg/ml of ampicillin at a ratio of 1%, and cultured at 37° C. for 6 hours. The 5 ml of the cultured broth was centrifuged to collect cells, which were then washed with a 0.85% NaCl solution. The cells were suspended in 3 ml of a reaction mixture containing 200 μ moles of an NH4Cl-NH4OH buffer (pH 9.0), 109 μ moles of a sodium phenylpyruvate, and 267 μ moles of a lactose, and the suspension was allowed to stand at 30° C. for 24 hours. As a result, 20 mg/ml (yield 33%) of L-phenylalanine was accumulated, as determined by the above-mentioned L-phenylalanine assay procedure.

Provenance & Evidence