Thiamine → L-Glutamic Acid

Participants

Reaction Steps

Both strains, VL334thrC+ and VL334thrC+-ΔcsrA, can be grown for 18-24 hours at 37° C. on L-agar plates. Then, one loop of the cells can be transferred into test tubes containing 2 ml of fermentation medium. The fermentation medium contains 60 g/l glucose, 25 g/l ammonium sulfate, 2 g/l KH2PO4, 1 g/l MgSO4, 0.1 mg/ml thiamine, 70 μg/ml L-isoleucine and 25 μl CaCO3 (pH 7.2). Glucose and CaCO3 should be sterilized separately. Cultivation can be carried out at 30° C. for 3 days with shaking. After the cultivation, the amount of L-glutamic acid produced can be determined by paper chromatography (liquid phase composition: butanol-acetic acid-water=4:1:1) with subsequent staining by ninhydrin (1% solution in acetone) and further elution of the compounds in 50% ethanol with 0.5% CdCl2.

Provenance & Evidence