O=C[C@@H]([C@H]([C@@H]([C@@H](CO)O)O)O)O.S([O-])([O-])(=O)=O.[NH4+].[NH4+].OP([O-])(O)=O.[K+].[O-]S([O-])(=O)=O.[Mg+2].CC1[N+](CC2C=NC(C)=NC=2N)=CSC=1CCO.[NH2:50][C@H:51]([C:56]([OH:58])=[O:57])[C@H:52](CC)[CH3:53].[C:59]([O-:62])([O-:61])=O.[Ca+2]>>[NH2:50][C@H:51]([C:56]([OH:58])=[O:57])[CH2:52][CH2:53][C:59]([OH:62])=[O:61]Both strains, VL334thrC+ and VL334thrC-Δtarget gene, 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 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). The pH is adjusted to 7.2. Glucose and CaCO3 are sterilized separately. Cultivation can be carried out at 30° C. for 3 days with shaking. After the cultivation, the amount of L-glutamic acid which is produced can be determined by paper chromatography (liquid phase composition of 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.
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