An efficient procedure for the production of L-[4-C-13]aspartic acid (4-C-1
3-Asp) was investigated. In this procedure, phosphoenolpyruvate carboxylase
originating from the methanol-assimilating microbe, Methylobacterium extor
quens JCM 2805, was used for the production of labeled oxaloacetic acid fro
m phosphoenolpyruvate (PEP) and (NaHCO3)-C-13; the oxaloacetic acid was the
n converted to 4-C-13-Asp with glutamic-oxaloacetic transaminase. In this r
eaction, with starting concentrations of 10 mM PEP, 10 mM (NaHCO3)-C-13 and
15 mM L-glutamic acid (Glu), the yield was 70%. 4-C-13-Asp and Glu in the
final reaction mixture were separated by displacement chromatography. The y
ield of this process was 84%. The overall yield was 59%. The incorporation
of C-13 at the C-4 position of 4-C-13-Asp was confirmed by NMR spectroscopy
.