Thiamine → L-Glutamic Acid

関与物質

反応手順

Both strains, VL334thrC+ and VL334thrC+-ΔglgC, 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 a fermentation medium. The fermentation medium (pH 7.2) should contain glucose (60 g/l), ammonium sulfate (25 g/l), KH2PO4 (2 g/l), MgSO4 (1 g/l), thiamine (0.1 mg/ml), L-isoleucine (70 μg/ml), and CaCO3 (25 g/l). 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.

情報源と証拠