Thiamine → L-Glutamic Acid

HRID 1190784Notes

Reaction equation

HRID 1190784 reaction equation
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.[NH2:38][C@H:39]([C:44]([OH:46])=[O:45])[C@H:40](CC)[CH3:41].[C:47]([O-:50])([O-:49])=O.[Ca+2]>C(O)C>[NH2:38][C@H:39]([C:44]([OH:46])=[O:45])[CH2:40][CH2:41][C:47]([OH:50])=[O:49]

Participants

8

Reactant 6

HCID 1130ThiamineHCID 6306L-IsoleucineHCID 10112Calcium Carbonate2×HCID 24083Sulfuric acid magnesium salt (1:1)HCID 516951Monobasic potassium phosphateHCID 6097028Ammonium Sulfate

Product 1

HCID 33032L-Glutamic Acid

Reagents, catalysts & solvents 1

HCID 702EthanolSolvent

Procedure

Both strains, VL334thrC+ Pl-tacadhE* and VL334thrC+ Pl-tacadhE* (pPLydbK3), 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 ethanol (20 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. CaCO3 is 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.

Workup

  1. additioncontaining 2 ml of fermentation medium
  2. waitCultivation can be carried out at 30° C. for 3 days

Sources & evidence

ORD record
ord-44ff7937dcf943cbae202fc374617c84
Source dataset
uspto-grants-2014_03
Patent number
US08679798B2

Reaction notes

Loading…